Adoptive cellular therapy of urological neoplastic disease

Cancer

Treatment

Reviews

(1994)

20,

173-l

90

ANTITUMOR TREATMENT Adoptive disease

cellular

Jay E. Gold,*

Michael

therapy

E. Osbandt

of urological

and Robert

neoplastic

J. KraneS

* The Divisions of Hematology and Neoplastic Diseases, Department of Medicine, The Mount Sinai Hospital and Mount Sinai School of Medicine of the City University of New York, New York, N.Y.; t The Division of Pediatric HematologyOncology, Department of Pediatrics and the *Department of Urology, Boston City Hospital and Boston University School of Medicine, Boston, MA.

Introduction Adoptive cellular therapy of neoplastic disease is the transfer of previously sensitized immune cells to tumor-bearing hosts (TBH) in order to achieve therapeutic effect and is based on experimental evidence that the cellular arm of the immune system is the crucial factor in mediating tumor rejection (1). One of the first examples of adoptive cellular therapy occurred more than 30 years ago, when Mitchison demonstrated that murine TBH treated with syngeneic immune cells could be cured of their disease (2). Later, Bach and others demonstrated the concept of poolpriming using lymphocytes from random human TBH with lymphoma or leukemia. When stimulated ex vivo by one or more sets of irradiated allogeneic peripheral blood mononuclear cells (PBMC) from tumor-free hosts, these TBH-derived lymphocytes were found to be effective tumor killers ex vivo (3, 4). Mazumder and co-workers reported that mitogen-activated lymphocytes, using phytohemagglutinin (PHA), were able to be activated ex vivo and then be safely infused into human TBH (5). Using newly described cytokines such as the interferons and interleukins, especially interleukin-2 (IL-2), it became possible to activate PBMC or splenocytes from murine and human TBH ex vivo. These IL-2-activated PBMC are known as lymphokine-activated killer (LAK)-cells (6-8). LAK-cells are able to eradicate, in a non-specific fashion, various syngeneic and allogeneic tumor targets but not syngeneic or autologous healthy tissue. In addition, other clinical adoptive cellular therapy trials in humans have included the use of antitumor specific cellular immune therapies including tumor-infiltrating lymphocytes (TIL), and autolymphocyte therapy (ALT). Many of the clinical studies of adoptive cellular therapy in huma.ns have dealt with the application of adoptive cell transfer for the treatment of metastatic renal Correspondence should be addressed to: Jay E. Gold, The of Medicine, 920 Park Avenue, New York, N.Y. 10028, U.S.A. Sponsored in part by a grant from The Jennifer Turner Cancer 0305-7372/94/020173+18

Mount Research

Sinai

Medical

Center,

Foundation. @ 1994

$08.00/O 173

Department

W.0.

Saunders

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J. E. GOLD

ETAL.

Mononuclear

cell

subsets

#+$#Q -

Addition of exogenous cytokines and/or MoAb[IL-2 t TNF, IFN, anti, -CDz, etcl

Figure 7. Illustration of LAK and TlL therapies. LAK-cells are derived from PBMC obtained via apheresis and are composed mostly of NK-cells (a). TIL are obtained from solid tumor lesions and are composed mostly of T-cells (b). Other cells contributing to LAK and TIL as shown are monocytes (M), B-cells (B), and other mononuclear cell populations (0). These cells are expanded and activated ex viva with IL-2 and/or as exogenous cytokines and/or monoclonal antibodies (MoAb). The activated cells are then re-infused into TBH with concomitant administration of exogenous IL-2.

cell carcinoma (RCC). Therefore, the bu!k of this review will focus on the of both non-tumor specific as well as tumor-specific adoptive cellular the treatment of metastatic RCC. In addition, methods to augment the potential of adoptive cellular therapy in the treatment of RCC as application to other urological tumors such as prostate carcinoma transitional cell carcinoma of the bladder (TCC) will also be addressed.

Adoptive

cellular

L ymphokine-activated

therapy

with

non-tumor

specific

application therapy to therapeutic well as its (PCA) and

lymphocytes

killer cells

PBMC obtained from TBH that are cultured short term in relatively high concentrations of IL-2 results in the generation of cells that are able to lyse fresh, noncultured, natural killer (NK) -cell resistant tumor cells, but not normal cells (6). Early reports described these LAK-cells as distinct from T, B, or NK-cells (see Figure l), as they were able to cause ex vivo lysis of NK-resistant targets such as the Daudicell line (7, 8). More recent reports confirm that most LAK-cells are of NK origin, with some contribution from T-cells and B-cells (6-8). Because the majority of LAK-cells are made up of NK-cells and lack T-cell receptors, they are able to kill both autologous and allogeneic tumor targets in a non-specific or non-major histocompatibility complex (MHC)-restricted manner (9, IO). The success of IL-2-based adoptive cellular therapy in animal models (11-14) prompted clinical trials of LAK-cells and IL-2 in human TBH by Rosenberg and coworkers at the National Cancer Institute (NCI) (15-I 7). Unfortunately, the success

ADOPTIVE Table

Author

I.

CELLULAR

THERAPY

Adoptive cellular therapy with LAK-cells and IL-2 (Ref.)

No. patients

OF GU CANCER of renal

cell

175

carcinoma

CR

PR

CR+PR

Rosenberg (17) Rosenberg (17a) West (23) Schoof (24) Thompson (26) Wang (67) Paciucci (27) Fisher (18) Parkinson (25) Sznoll (114)

54 48 6 10 8 32 9 32 47 40

7 7 0 0 1 2 0 2 2 0

10 8 3 5 0 5 1 3 2 8

17 15 3 5 1 7 1 5 4 8

Totals CR = complete response PR = partial response.

286

21

45

66(23%)

of the murine LAK/IL-2 experiments could not be duplicated to the same degree in the human triais. The initial studies from Rosenberg’s group have been updated and some responses in patients with various malignancies have been noted, especially in those with melanoma and RCC (Table 1) (15-17a). Responses in RCC were also noted by Fisher and colleagues in the extramural LAK/IL-2 study although at a lower frequency than the NCI data (18). Five of 32 patients in the extramural study achieved a response that included two complete responses and three partial responses. Of note is that in two of the three patients with a partial response, surgical resection of residual masses rendered them disease-free. It should also be pointed out that the patients in the extramural trials had poorer prognostic features than those in the NCI trial such as residual primary tumors and/or large tumor masses, which may explain the lower response rates observed. The results of other LAK/IL-2 trials in RCC are also shown in Table 1, with response rates consistent with the NCI and Extramural trials. Based on these data, it appears that adoptive cellular therapy using LAK/IL-2 is most effective in patients with low tumor burdens as well as those patients whose primary tumor has been removed, an observation that has been confirmed in murine models (19). Because of the relatively low response rate, the severe toxicities (20-22) associated with IL-2-dependent adoptive cellular therapy (Table 2) and the prohibitive cost per patient, it is unlikely that LAK/IL-2 in its current state would be effective as a widespread form of adoptive cellular therapy. Methods to attenuate the treatment toxicity, such as continuous venous infusion (CVI) or bolus injection (BI), with lower doses than the original NCI/Extramural trials, have been attempted (23-27). The toxicity of CVI vs. BI appears significantly less, but this may be at the expense of a lower complete response rate in the CVI patients as they had received a lower total IL-2 dose than the BI patients. Other methods to decrease toxicity have employed local or intralymphatic infusion of LAK/IL-2, both in animal models and in a small number of human trials (28-30). The “bi-compartmental” model of Wiltrout and co-workers for metastatic murine RCC employed both systemic and local therapy with LAK/IL-2 for primary and metastatic disease (31, 32). Further studies will have to delineate the utility of LAK/IL-2 in adoptive cellular therapy and whether non-tumor specific LAK/IL-2 would be more advantageous than tumor-specific adoptive cellular therapy.

176

J. E. GOLD Table l

l

l

l

0

l

0

l

2.

Toxicities

associated

with

ETAL.

IL-2-dependent

adoptive

cellular

therapy

Capillary leak syndrome Fluid retention Hypotension Pulmonary edema Cardiac Arrythmias Myocardial depression Myocardial infarction Renal Oliguria Prerenal azotemia Neuropsychiatric Somnolence, lethargy Behavioral changes Cognitive changes Hepatic Cholestasis Hyperbilirubinemia Gastrointestinal Diarrhea, nausea, vomiting Stomatitis Bowel perforation Hematologic Anemia Thrombocytopenia Eosinophilia Neutrophil chemotactic defect Dermatologic Macular erythema Desquamation Pruritis

Adoptive Tumor-infiltrating

cellular

therapy

with

tumor-specific

T-cells

lymphocytes

It is well known that mononuclear cells can be found infiltrating various solid neoplasms (33-40). Most of these tumor-infiltrating lymphocytes (TIL) have been identified as either CD4’ (helper/inducer) or CD8’ (cytotoxic/suppressor) T-cells. Some mononuclear cells are also of NK-cell lineage and bear NK markers such as CD56 (LeulS/NKH-1) or CD57 (Leu7/HNK-1). One can also find macrophages and B-cells in the mononuclear cell infiltrate depending on the tumor type. The nature and etiology of this mononuclear cell infiltrate is somewhat controversial, although many believe that it signals a host immunologic response to the tumor, which in turn may indicate a favorable response for the patient. Yet, should this be the case, questions persist as to why these mononuclear cells do not eradicate the primary tumor and why tumor dissemination can still occur despite a ‘favorable’ immune response. In addition, TIL proliferate and kill tumor targets poorly when isolated from tumors (41). It is possible that TIL are either immune-suppressed from the tumor milieu or that suppressor cells themselves make up a significant fraction of the TIL and hence contribute to the ineffectiveness of these TIL in attempting to eradicate the tumor (42-45). The basic principles of harvesting, isolating, and expanding TIL for adoptive cellular therapy from solid tumor lesions is illustrated in Figure 1. Rosenberg’s group

ADOPTIVE Table

Author

3.

Adoptive with TIL

CELLULAR cellular and IL-2

(Ref.)

Topalian (53) Kradin (52) Oldham (50) Hanson (115) Belldegrun (116) Hanson (50a) Totals CR = complete response PR = partial response. SD = stable disease.

THERAPY therapy

No. patients 4 7 9

OF GU CANCER of

renal

cell

177

carcinoma

Response

:‘5 14

1 (PR) 2 (PR) 3 (SD) 5 (SD) 8 (5CR.3PR) 10 (1 PR,SSD)

71

29 (41%)

noted that when small minced or enzyme-digested fragments of murine tumors containing TIL were incubated in IL-2, the TIL expanded and proliferated in culture, causing destruction of the tumor (46). When these IL-2-activated TIL were given to murine TBH treated with cyclophosphamide (CY), a large number had their tumors eradicated. TIL were also noted to be 50-100 times more potent tumor killers on a per cell basis than LAK-cells. However, when human TIL were expanded with IL-2, many of these TIL, depending on the tumor type, appear to lose their MHC-restricted tumor specificity and become ‘LAK-like’ in their killing of nonautologous tumor targets (47). Indeed, studies of IL-2-generated TIL in patients with RCC, PCA, or TCC demonstrate enhanced ex vivo tumor killing of TIL vs LAKcells, although the killing was not restricted to autologous tumors since allogeneic RCC, PCA, and TCC targets as well as Daudi and K562 cell lines were also lysed (35, 36). As mentioned, some of these TIL consist of CD3’CD56’ T-cells as well as CD3-CD16’CD56’ NK-cells, implying that many of these mononuclear cells may be non-MHC-restricted in their tumor killing. ltoh and Balch demonstrated that TIL from melanoma patients possess autologous tumor specificity after IL-2 expansion (39), an observation shared by Kradin and Kurnick who studied TIL from lung cancer patients (48, 48a). However, Maleckar and co-workers using their tumor-derived activated cells (TDAC), demonstrated that these TDAC could be made relatively, as opposed to absolutely, tumor specific by frequent re-stimulation with irradiated, autologous tumor cells (49, 50). Finke and Bukowski found that repetitive re-stimulation of RCC TIL with autologous tumor generated CD4’ T-cells that were generally not restricted to autologous RCC targets (51). It seems that the putative tumor specificity of TIL may vary by tumor type or may be a relative phenomenon in all tumor types. The responses to TIL/IL-2 therapy in humans are shown in Table 3. Kradin and co-workers demonstrated partial and minor responses in 43% of patients with RCC (52). The study by Topalian’s group obtained a partial response in one of four patients with metastatic RCC (53). Hanson demonstrated responses in five of 12 patients and Belldegrun noted eight responses (five CR, three PR) in 25 patients (32%) with metastatic RCC (115, 116). Oldham and co-workers treated nine patients with metastatic RCC using TDAC/IL-2. Although no objective responses were seen, three patients did demonstrate stable disease (50). Hanson’s group used TIL/IL-2 in 14 patients with metastatic RCC and demonstrated a partial response in one patient and stable disease in nine patients (50a).

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ETAL.

The toxicity of the TIL/IL-2 trials was substantial and similar to previous LAK/IL2 studies. In a similar fashion to the LAK/IL-2 trials, it appears that the best responses are obtained by bolus dosing, despite greater toxicity. Overall, therapy with TIL/IL-2 appeared to show better antitumor responses in metastatic RCC than LAK/IL-2. As with LAK/IL-2, the tumor burden in these patients may play a role in patient refractoriness to TIL/IL-2. In addition, the ability of TIL to accumulate at tumor sites may play a role in the degree of antitumor responses seen. Reports by Muhkerji and Rosenberg have shown that indium-labelled TIL do indeed traffic to tumor sites (54, 55). Another possible explanation for lack of greater antitumor responses is that IL-2 generated TIL may contain a significant number of suppressor cells, which would impair their effectiveness in adoptive cellular therapy (42). Since IL-2-expanded TIL contain both CD4’ and CD8’ T-cells, the question arises as to which TIL subpopulation might contain the putative suppressor cells. Muhkerji’s group demonstrated that cloned PBMC CD4’ T-cells that mediated autologous tumor killing had their killing ability severely reduced in the presence of CD8’ Tlymph node lymphocytes (LNL) from the tumor-invaded lymph nodes but not by CD4’ T-LNL in a patient with a metastatic paraganglionoma (43). However, these same investigators noted that both CD4’ T-LNL and CD8’ T-LNL from patients with metastatic melanoma, inhibited the ex viva generation of PBMC T-cell killing in a specific fashion against the autologous melanoma, implying that different tumors may induce different subset(s) of suppressor T-cells (44, 45). A recent report by Finke and co-workers may explain the functional basis of these impaired TIL (45a). They demonstrated that TIL T-cells rather than peripheral blood T-cells from TBH with RCC may have impaired antitumor responses secondary to decreased expression of the T-cell receptor [ chain and ~56’~~ tyrosine kinase. The generation of TIL is much more labor-intensive than LAK cells, requiring a median time of 8 weeks to propagate in the laboratory (56). In addition, TIL from some patients fail to grow in culture despite optimal conditions. TIL require restimulation during these long-term cultures to maintain their tumor killing ability (48, 51). Despite these limitations, TIL therapy is not without therapeutic potential. TIL have also been isolated and expanded ex viva from non-RCC solid tumors that include PCA and TCC (35, 36). The ability to expand TIL ex viva from tumors may indicate an active antitumor T-cell surveillance and hence give a better overall prognosis and clinical response than those patients treated with LAK/IL-2. At this point, considering its possible tumor-specificity and clinical superiority, there may be an advantage in TIL-based adoptive cellular therapy for urological neoplastic disease using relatively tumor-specific T-cell therapy with TIL/IL-2 rather than LAK/IL-2. Autolymphocyte

therapy

Autolymphocyte therapy (ALT) is outpatient autologous adoptive cellular therapy that is not dependent on exogenous IL-2. PBMC obtained from TBH are activated ex vivo using an autologous lymphokine mixture (T3CS), which in turn is derived from the culture supernatant following incubation of PBMC from TBH in low doses of the mitogenic anti-CD3 monoclonal antibody (MoAb), OKT3. T3CS-activated PBMC (termed autolymphocytes or ALT-cells) are then re-infused into the patients with a minimum of side-effects (57-60). As reported in murine tumor models and

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Figure 2. Illustration of ALT. PBMC obtained from TBH (1) are incubated ex viva with low doses of the mitogenic MoAb, OKT3 (2). The supernatant derived from this process (T3CS) is then combined (3) with autologous PBL obtained from TBH (4) resulting in the generation of ALT-cells (5). These ALTcells are then re-infused into TBH without the use of exogenous IL-2 (6).

then human TBH, ALT-cells are composed primarily of antitumor-specific CD45RO’ (human) or CD44’ (mouse) memory T-cells that have been non-specifically activated ex vivo via T3CS (61-63). Analysis of T3CS has shown it to contain small amounts of OKT3 as well as being rich in cytokines such as GM-CSF, IFN-y, IL-l, and IL-6, although not IL-2 (121). It appears that it is possible to generate tumorspecific recall responses following stimulation with anti-CD3 antibodies. This was also confirmed by Cheever and Greenberg who demonstrated that spleen cells obtained from mice with FBL-3 erythroleukemia and cultured with anti-CD3 antibody, demonstrated preferential tumor-specific proliferative and cytolytic activity when stimulated with FBL-3 cells following anti-CD3 activation (64). To reduce tumor-associated suppressor cell activity in these TBH, oral cimetidine is given as part of the ALT protocol as suppressor T-cells have been shown to express H;, receptors on their cell surfaces. In a preliminary randomized trial, patients with metastatic RCC who were treated with ALT had a 3-4 fold longer survival with improved quality of life compared to those in the control group (65). A recent update of the initial study, as well as a new study with additional patients, confirmed both the survival advantage and response rate of 18% in the ALT-treated group (66). These results seem to indicate that prolonged survival with improved quality of life in a significant number of patients treated with virtually toxicity-free adoptive cellular therapy may be more effective than therapies where few complete or transient partial responses were obtained, a fair amount of toxicity occurred and no real impact on survival was observed. However, it must be emphasized that the ALT responses (or lack, thereof) were judged radiographically. It may be that a number of these patients were indeed

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responders, but with residual, non-viable tumor masses seen on x-ray studies. Surgical resection, as done in the study by Fisher and co-workers, may convert these patients into radiographic complete responders (18). Indeed, a patient with documented metastatic RCC to the liver who underwent ALT was found to have fibrotic, non-viable tumor masses during an emergency cholecystectomy (Osband M.E., personal communication). Asimilar conclusion regarding maintenance therapy and prolonged survival was reached by Wang and co-workers in their study using periodate-activated cells and IL-2 (67). They showed that while the complete response rate was not significantly affected, patients had a greater survival with maintenance therapy. This concept of prolonged survival by immunotherapy was also demonstrated by lkarashi and co-workers following adoptive transfer of tumorinfiltrating lymphocytes in patients with ovarian cancer (67a). Other correlations with prolonged survival in ALT-treated patients have been noted (65, 68, 69). Serum markers that have correlated with survival in ALT treatment are soluble IL-2 receptor (IL-2R) and immunosuppressive acidic protein (IAP) (68). High levels of both IL-2R and IAP in patients with metastatic RCC conferred a negative survival advantage. When the phenotype of the T-cells in the ALT-cell preparation were examined following ex viva modulation, RCC patients whose T-cells predominantly expressed IL-2R’ and la’ had a lower survival rate compared to those whose T-cells predominantly expressed the VLA-1 marker (67, 68). In the adoptive cellular therapy trials using LAK and TIL it had been thought that the greater number of antitumor cells infused, the greater the antitumor response. However, a study by Ross and Osband on patients treated with ALT found a bimodal distribution of infused cell number with a significant impact on survival (72). They found that above and below a certain critical number of cells per infusion, patients actually had a decreased survival. It is possible that a certain number of cells provokes an effective antitumor response and once that critical number of cells has been exceeded, one may provoke an anti-idiotypic, anti-ALT response downregulating and suppressing antitumor effectiveness. In addition to ALT, this may also have significance in LAK and TIL therapies.

Future

directions

The use of antitumor immune cells as therapy for malignant disease represents a potentially valuable potential tool in addition to the traditional tools of surgery, radiotherapy, and chemotherapy in urologic oncology. Adoptively transferred ex viva activated lymphocytes have demonstrated activity in two diseases (RCC and melanoma) for which other treatment modalities are generally ineffective. The task now at hand is to improve upon what is currently available by enhancing antitumor effectiveness and decreasing toxicity. It appears that among potentially effective antitumor immune cells, T-cells with tumor-specific immunity are likely to be the most potent. However, human cancers are much less immunogeneic than their animal counterparts and so may make the isolation and expansion of tumor-specific T-cells problematic. Additionally, it has been demonstrated that human tumors can down-regulate their cell surface MHC

ADOPTIVE

CELLULAR

THERAPY

OF GU CANCER

181

expression, possibly as a selection effect against a cytotoxic T-cell response, making them less immunogenic and therefore potentially less recognizable to antitumorspecific T-cells (120). Approaches to convert poorly immunogenic tumors into immunogenic ones include xenogenization with chemotherapy drugs and viruses, as well as transduction of genes encoding antigens into tumor cells (73-76). Townsend and Allison have shown that human melanoma cells transfected with the CD28 co-stimulatory ligand B7, are able to be lysed by CD8’ T-cells (77a). While TIL therapy apparently holds the promise of adoptive T-cell-based antitumor-specific cellular therapy, difficulties in obtaining and expanding sufficient numbers of cells for use is a major limitation to the application of this form of adoptive cellular therapy on a widespread basis. In addition, not all patients have tumor tissue available for biopsy and true tumor-specificity appears restricted to melanoma only. Methods to circumvent these limitations include ex vivo stimulation of TIL using anti-CD3/TCR antibodies (77-81). Using these antibodies, it is possible to generate adequate numbers of antitumor T-cells with increased cytotoxicity for adoptive transfer in pre-clinical studies with both LAK-cells and TIL. In addition, it is this methodology that forms the basis for ALT where memory T-cells are generated ex vivo using the supernatant (T3CS) derived from PBMC stimulated with antiCD3 antibodies. As a further illustration of the efficacy of anti-CD3-based activation, Chang and Shu have also demonstrated that T-cells obtained from lymph nodes draining established tumors are also capable of specific antitumor activity following activation by anti-CD3 antibodies (82). These in vitro sensitized or IVS T-cells have been shown to be effective in murine sarcoma models and trials are currently underway in humans (83). Additionally, Cheever hasshown in murine hematopoietic tumors that splenocytes from FBL-3 leukemia-bearing mice can cure syngeneic leukemia-bearing mice following ex vivo activation with anti-CD3 antibodies (64). Taking this a step further, Kradin and co-workers used bi-specific anti-CD3 antibodies selectively to expand CD4’ or CD8’ T-cell subsets of TIL which may prove more therapeutically effective than both sets combined (84). This may also help remove any cells that are functioning as suppressors. In addition to determining better methods of antitumor T-cell isolation and expansion, it may be possible to direct these cells to their targets in vivo. Using hybrid antibodies consisting of CD3 and monoclonal antibodies against a particular tumor target, it has been shown that it is possible to focus T-cells directly onto a tumor cell (85, 86). Once at the tumor site, application of molecular biologic techniques may allow for the generation of more potent antitumor effector cells. Along these lines, it has been demonstrated that some TIL possess clonal rearrangements of their T-cell receptor V, genes (87). This may imply specificity to some tumor-associated antigen (TAA), although it is not clear whether cells with this rearrangement are functioning as helper or suppressor cells (88). Isolation and identification of the properties of these clonal cells as done in the murine 3LL tumor model by Gelber and co-workers may prove invaluable in future therapies (119). Using gene transduction therapy, Rosenberg’s group introduced the neomycinresistance gene into TIL and was able to isolate those TIL containing the gene from tumor sites (89, 90). This demonstrates that transduction of TIL with specific cytokine genes such as those encoding TNF or IFN may be able to be inserted into the genome and deliver a greater lethal effect at the tumor site. Additionally, ex vivo cytokine gene insertion of either IL-2 or IL-4 into tumor cells themselves has

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been shown to cause accumulation of antitumor effector cells at the tumor site resulting in destruction of the tumor (91, 92). Augmentation of adoptive cellular therapy may be possible through combinations of adoptively transferred T-cells with either chemotherapy [adoptive chemoimmunotherapy, (ACIT)] or radiation therapy (xRT). Previous studies have demonstrated that, in experimental disseminated animal solid and hematopoietic tumors which could not be cured by adoptive cellular therapy alone, cure was possible by either ACIT or adoptive cellular therapy plus xRT (93-95). Pre-clinical studies using adoptive cellular therapy in combination with low dose xRT have shown that the low dose xRT causes selective localization of antitumor effector cells at tumor sites (96). In the bi-compartmental model of Salup and Wiltrout, mice with LAK-cellresistant RCC were cured only when systemic chemotherapy using doxorubicin (DOX) was added to the LAK/IL-2 therapy (31, 32). It is postulated that aside from direct antitumor effects, which may aid in reducing the tumor burden, xRT and many chemotherapeutic agents have immunomodulatory properties (97). Gold and co-workers, using ALT in combination with CY, demonstrated cfinical efficacy in patients with relapsed but not refractory solid tumors (98). Similarly, melanoma patients from the NCI TIL/IL-2 trial also received ACIT with CY as the chemotherapy agent (99). Sznoll’s group combined DOX and CY with LAK/IL-2 in a variety of human solid tumors (114). There were eight partial responses seen in 40 evaluable patients with metastatic RCC. The study by Hanson and co-workers combined CY with TI L/I L-2 (50a). Formelli’s group demonstrated synergy of adoptively transferred T-cells and cis-diamminedichloroplatinum(ll) (CDDP) in a mouse lymphoma model (100). Studies by Gold and Mizutani showed markedly enhanced ex viva killing of human RCC and TCC tumor targets incubated in non-tumoricidal concentrations of CDDP by ALT-cells and LAK-cells, respectively (67, 101). As the dose of CDDP employed had no effect on tumor target viability, it appears that the synergistic antitumor effect was secondary to immunomodulation of the tumor targets. In addition to synergy with chemotherapeutic agents, potentiation of adoptive cellular therapy may be accomplished through combinations of cells with exogenous cytokines. LAK-cells, TIL, and IVS T-cells have shown greater antitumor efficacy ex viva and in murine TBH when used together with cytokines other than IL-2 (70, 102-104, 117, 118). In humans, Sznoll’s group combined LAK/IL-2 DOX, and CY with a-IFN in their treatment of metastatic RCC patients with results as previously described. Regarding ALT, patients whose T3CS demonstrated increased levels of IL-l had a sixfold survival advantage over those patients whose T3CS did not have increased IL-l levels (65). It has been demonstrated that IL-l as well as IL-6 in the T3CS causes the preferential expansion of CD45RO’ T-cells (63). In addition, other studies confirm that IL-l may play a significant role in adoptive cellular therapy using LAK-cells (70). Therefore, the addition of chemotherapy, xRT, or cytokines to adoptive cellular therapy may lead to more effective immunotherapy as mechanisms of synergy in these combinations become elucidated. Aside from LAK, TIL, or ALT, other tumor-immune lymphocytes may prove useful for adoptive cellular therapy. Adherent-Lymphokine-Activated Killer (A-LAK)-cells were discovered when it was found that certain MNC from PBMC and tumor specimens adhere to plastic surfaces such as culture flasks (106). A-LAK-cells are activated ex viva with IL-2 and are composed mostly of cells with an NKlike CD3-CD56’ phenotype, although CD3’CD56’ A-LAK-cells have also been

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described. A-LAK-cells appear to be more potent tumor killers ex viva than LAKcells and so may lead to more effective adoptive cellular therapy in human TBH when combined with IL-2. As previously mentioned, the IVS T-cells of Chang and Shu have been shown to be effective adoptive cellular therapy in murine tumor models, with studies in human TBH now underway. Unlike animal models of neoplasia, antigens that are truly tumor-specific have traditionally been difficult to demonstrate in human beings. However, recent evidence points to the existence of tumor-associated antigens (TAA) in humans. Such TAA include the Thomsen-Freidenreich (TF, Tn) antigens found on all epithelial human cancers as well as TCC and PCA (107-109). Another candidate TAA is prostate-specific antigen (PSA) as it is expressed on prostate tissue only (110). Osband and co-workers have succeeded in immunizing CD45RO’ (memory) T-cells against PSA ex vivo and demonstrated safety and feasibility of infusion of these anti-PSA T-cells in patients with metastatic PCA (113). Longenecker and co-workers have shown that synthetic compounds derived from TF or Tn are capable of delayed hypersensitivity responses when given as a therapeutic vaccine. CY plus vaccine in mice-bearing a lethal mammary carcinoma are capable of prolonging survival and curing tumor-bearing mice (111,112). In human TBH, Shu and Chang demonstrated improved clinical responses in patients receiving adoptively transferred IVS T-cells using ex vivo activation with a tumor vaccine (122). We have summarized the current status of adoptive cellular therapy in urological neoplastic disease. In particular, the manipulation of T-cells ex vivo with adoptive transfer of T-cell immunity to TBH with weakly immunogenic tumors, as well as methods to increase immunogenicity of tumor targets holds the promise for the potential success of adoptive cellular therapy in patients with advanced metastatic genitourinary tumors.

Acknowledgments The authors wish to thank Harris M. Nagler, Nadine Medley, and Steven H. Itzkowitz, for their continued support as well as for their comments and critique of the manuscript.

References 1. Borberg, H., Oettgen, H. F., Choudry, K. & Beattie E. J., Jr (1972) Inhibition of established transplants of chemically induced sarcomas in syngeneic mice by lymphocytes from immunized donors. ht. J. Cancer 10: 539-545. 2. Mitchison, N. A. (1957) Adaptive transfer of immune reactions by cells. J. Cell. Compar. fhys. 50: 247-254. 3. Zarling, J. M. & Bach, F. H. (1979) Continuous culture of T cells cytotoxic for autologous human leukaemia cells. Nature 280: 685-688. 4. Bach, M. L., Bach, F. H. & Zarling, J. M. (1978) Pool-priming: A means of generating T lymphocytes cytotoxic to tumour or virus-infected cells. Lancet l(8054): 20-22. 5. Mazumder, A., Eberlein, T. J., Grimm, E. A., Wilson, D. J., Keenan, A. M., Aamodt, R. & Rosenberg, S. A. (1984) Phase I study of the adoptive immunotherapy of human cancer with lectin activated autologous mononuclear cells. Cancer 53: 8964391.

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J. E. GOLD

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6. Grimm, E. A., Mazumder, A., Zhang, H. Z. & Rosenberg S. A. (1982) The lymphokine activated killer cell phenomenon: lysis of NK resistant fresh solid tumor cells by IL-2 activated autologous human peripheral blood lymphocytes. J. Exp. Med. 155: 1823-I 832. 7. Phillips, J. H. & Lanier, L. L. (1986) Dissection of the LAK phenomenon: relative contribution of peripheral blood NK cells and T lymphocytes to cytolysis. J. Exp. Med. 164: 314-324. 8. Damle, N. K., Doyle, L. V. & Bradley, E. C. (1986) IL-2 activated human killer cells as derived from phenotypically heterogenous precursors. J. lmmunol. 137: 28142821. 9. Zocci, M. R., Bottino, C., Ferrini, S., Moretta, L. & Moretta, A. (1987) A novel 120 KD surface antigen expressed by a subset of human lymphocytes. Evidence that lymphokine activated killer cells express this molecule and use it in their effector function. J. Exp. Med. 166: 319-331. 10. Chen, B. P., Malkovsky, M., Hank, J. A., Molendu, J. A. & Sondel, P. M. (1987) Non-MHCrestricted cytotoxicity (NRC) of normal peripheral blood mononuclear leukocytes (PBM) by PBM expanded with interleukin-2: NRC is non-specific and inhibited by monoclonal antibodies (MoAb) to LFA-1 but enhanced by MoAb to CD3 and HCA class I molecules. Cell. Immunol. 110: 382393. 11. Mule, J. J., Shu, S. & Rosenberg, S. A. (1982) The antitumor efficacy of lymphokine activated killer cells and recombinant IL-2 in vitro. J. lmmunol. 135: 646-653. 12. Cheever, M. A., Greenberg, P. D., Fefer, A. & Gillis, S. (1985) Augmentation of the antitumor therapeutic efficacy of long-term cultured T lymphocytes by in vitro administration of purified interleukin-2. J. Exp. Med. 155: 968-979. 13. Lafreniere, R. & Rosenberg, S. A. (1985) Successful immunotherapy of murine experimental metastases with lymphokine-activated killer cells and recombinant interleukin-2. Cancer Res 45: 3735-3742. 14. Pap, M. Z., Mule, J. J. & Rosenberg, S. A. (1986) Antitumor efficacy of lymphokine-activated killer cells and recombinant interleukin-2 in vivo: Successful immunotherapy of established pulmonary metastases from weakly immunogenic and nonimmunogenic murine tumors of three distinct histological types. Cancer Res. 46: 49734981. 15. Rosenberg, S. A., Lotze, M. T., Muul, L. M., Leitman, S., Chang, A. E., Ettinghausen, S. E., Matory, Y. L., Skibber, J. M., Shiloni, E., Vetto, J. T., Seipp, C. A., Simpson, C. & Reichert, C. M. (1985) Observations on the systemic administration of autologous lymhokine-activated killer cells and recombinant interleukin-2 to patients with metastatic cancer. N. Engl. J. Med. 313: 1485-l 492. 16. Rosenberg, S. A., Lotze, M. T., Muul, L. M., Chang, A. E., Avis, F. P., Leitman, S., Linehan, W. M., Robertsen C. N., Lee, R. E., Rubin, J. T., Seipp, C. A., Simpson, C. G. & White, D. E. (1987) A progress report on the treatment of 157 patients with advanced cancer using lymphokine-activated killer cells and interleukin-2 or high-dose interleukin-2 alone. N. Engl. J. Med. 316: 889-897. 17. Rosenberg, S. A. (1988) The development of new immunotherapies for the treatment of cancer using interleukin-2. Ann. Surg. 206: 121-l 30. 17a. Rosenberg, S. A., Lotze, M. T., Yang, J. C., Topalian, S. L., Chang, A. E., Schwartentruber, D. J., Muul, L. M., Seipp, C. A., Simpson, C. G. & White, D. E. (1993) Prospective randomized trial of high-dose interleukin-2 alone or in conjunction with lymphokine-activated killer cells for the treatment of patients with advanced cancer. J. Nat/ Cancer Inst. 85: 622-632. 18. Fisher, R. I., Coltman, C. A., Doroshow, J. H., Rayner, A. A., Hawkins, M. J., Mier, J. W., Wiernik, P., McMannis, J. D., Weiss, G. R., Margolin, K. A., Gemlo, B. T., Hoth, D. F., Parkinson, D. R. & Paietta, E. (1988) Metastatic renal cancer treated with interleukin-2 and lymphokine-activated killer cells. A phase II clinical trial. Ann. Intern. Med. 108: 518-523. 19. Thompson, J. A., Peace, D. J., Klan-ret, J. P., Kern, D. E., Greenberg, P. D. & Cheever, M. A. (1985) Eradication of disseminated murine leukemia by treatment with high-dose interleukin-2. J. lmmunol. 137: 3675-3683. 20. Margolin, K. A., Rayner, A. A., Hawkins, M. J., Atkins, M. B., Dutcher, J. P., Fisher, R. I., Mier, J. W., Wiernik, P., McMannis, J. 0.. Weiss, G. R., Gemlo, B. T., Hoth, D. F., Parkinson, D. R. & Paietta, E. (1989) Interleukin-2 and lymphokine-activated killer cell therapy of solid tumors: Analysis of toxicity and management guidelines. J. C/in. &co/. 7: 486-492. 21. Herberman, R. B. (1989) Interleukin-2 therapy of human cancer: Potential benefits versus toxicity. J. Clin. Oncol. 7: 14. 22. Klempner, M. S., Noring, R., Mier, J. W. & Atkins M. B. (1990) An acquired chemotactic defect in neutrophils from patients receiving interleukin-2 immunotherapy. N. Engl. J. Med. 322: 959-964. 23. West, W. H., Taner, K. W., Yannelli, J. R., Marshall, G. D., Orr, D. W., Thurman, G. B. & Oldham,

ADOPTIVE

24.

25.

26.

27.

28. 29.

30.

31. 32. 33. 34.

35. 36. 37. 38.

39. 40.

41.

42. 43.

44.

CELLULAR

THERAPY

OF GU CANCER

185

R. K. (1987) Constant infusion recombinant interleukin-2 in adoptive immunotherapy of cancer. N. Engl. J. Med. 316: 898-904. Schoof, D. D., Gramolini, B. A., Davidson, 0. L., Massaro, A. F., Wilson, R. E. & Eberlein, T. J. (1988) Adoptive immunotherapy of human cancer using low-dose recombinant interleukin 2 and lymphokine-activated killer cells. Cancer Res. 48: 5007-5014. Parkinson, D. R., Fisher, R. I., Rayner, A. A., Paietta, E., Margolin, K. A., Weiss, G. R., Hawkins, M. J., Atkins, M. B., Dutcher, J. P., Mier, J. W., Wiernik, P., McMannis, J. D., Gemlo, B. T., Hoth, D. F. 81 Paietta, E. (1990) Therapy of renal cell carcinoma with interleukin-2 and lymphokine-activated killer cells: Phase II experience with a hybrid bolus and continuous infusion interleukin-2 regimen. J. Clin. Oncol. 8: 1630-l 637. Thompson, J. A., Lee, D. J. & Lindgren, C. G. (1989) Influence of schedule of interleukin-2 administration on therapy with interleukin-2 and lymphokine-activated killer cells. Cancer Res. 49: 235242. Paciucci, P. A., Holland, J. F., Glidewell, 0. & Odchimar, R. (1989) Recombinant interleukin-2 by continuous infusion and adoptive transfer of recombinant interleukin-2-activated cells in patients with advanced cancer. J. C/in. Oncol. 7: 869-877. Bubenik, J. (1990) Local and regional immunotherapy of cancer with interleukin-2. J. Cancer Res. Clin. Oncol. 116: 1-7. Forni, G., Cavallo, G. P., Giovareli, M., Benetton. G., Jemma, C. & Baroglio, M. G. (1988) Tumor immunotherapy by local injection of interleukin-2 and non-reactive lymphocytes. Prog. Exp. Tumor. Res. 32: 187-l 93. Yasumoto, K., Nagashima, A., Ishida, T., Kuda, T., Yeno, T. & Sugimachi, K. (1987) Induction of lymphokine activated killer cells by intrapleural instillations of recombinant interleukin-2 in patients with malignant pleurisy due to lung cancer. Cancer Res. 47: 2184-2190. Salup, R. R., Back, T. C. & Wiltrout, R. H. (1987) Successful treatment of advanced murine renal cell cancer by bicompartmental adoptive chemoimmunotherapy. J. lmmunol. 138: 641-649. Wiltrout, R. H. 81 Salup, R. R. (1988) Adoptive immunotherapy combination with chemotherapy for cancer treatment. Prog. Exp. Tumor. Res. 32: 128-l 35. Vose, B. M. 81 Moore, M. (1985) Human tumor-infiltrating lymphcytes: a marker of host response. Semin. Hematol. 22: 27-38. Balch, C. M., Riley, L. B., Bae, Y. J., Salmeron, M. A., Platsoucas, C. D. & von-Eshenbach, A. (1990) Patterns of human tumor-infiltrating lymphocytes in 120 human cancers. A&. Surg. 125: 200-208. Haas, G. P., Solomon, D. & Rosenberg, S. A. (1990) Tumor-infiltrating lymphocytes from nonrenal urological malignancies. Cancer Immunol. Immunother. 30: 342-347. Tsujihashi, H., Uejima, S., Akiyama, T. & Kurita, T. (1989) lmmunohistochemical detection of tissue-infiltrating lymphocytes in bladder tumors. U-o/. Int. 44: 5-l 3. Finke, J. H., Tubbs, R., Connelly, B., Pontes, E. & Montie, J. (1988) Tumor-infiltrating lymphocytes in patients with renal cell carcinoma. Ann. N.Y. Acad. Sci. 532: 387-933. Belldegrun, A., Muul, L. M. & Rosenberg, S. A. (1988) Interleukin-2 expanded tumor-infiltrating lymphocytes in human renal cell cancer: isolation, characterization and antitumor activity. Cancer Res. 46: 206-l 23. Itoh, K., Platsoucas, C. D., & Balch, C. M. (1988) Autologous tumor-specific cytotoxic T lymphocytes in the infiltrate of human metastatic melanomas. J. Exp. Med. 168: 1419-I 430. Belldegrun, A. & Rosenberg, S. A. (1987) Antitumor and proliferative responses of IL-2-expanded tumor infiltrating lymphcytes (TIL) vs lymphokine activated killer (LAK) cells derived from patients with advanced renal cell cancer (RCC). Fed. Proc. 46: 1508 (abstr). Miescher, S., Whiteside. T. L., Carrel, S. von Fliedner, V. (1986) Functional properties of tumorinfiltrating and blood lymphocytes in patients with solid tumors: Effects of tumor cells and their supernatants on proliferative responses of lymphocytes. J. lmmunol. 136: 1899-I 907. Mukherji, B., Ergin, M. T., Guha, A., Tatake, R. T., & Nashed, A. L. (1986) Suppressor cell activities in autologous human tumor systems. Arch. Surg. 121: 14041141. Mukherji, B., Guha, A., Loomis, R. & Ergin, M. T. (1987) Cell-mediated amplification and down regulation of cytotoxic immune response against autologous human cancer. J. Immunol. 138: 1987-l 994. Mukherji, B., Wilhelm, S. A., Guha, A. & Ergin, M. T. (1986) Regulation of cellular immune response against autologous human melanoma. I. Evidence for cell-mediated suppression of in vitro cytotoxic immune response. J. lmmunol. 136: 1888-l 997.

186 45.

45a.

46. 47.

48.

48a.

49.

50.

50a.

51.

52.

53.

54.

55.

56. 57.

58. 59. 60.

61.

J. E. GOLD

ET AL.

Mukherji, B., Nashed, A. L., Guha, A., & Ergin, M. T. (1986) Regulation of cellular immune response against autologous human melanoma. II. Mechanism of induction and specificity of suppression. J. lmmunol. 136: 1893-l 901. Finke, J. H., Zea, A. H., Stanley, J., Longo, D. L., Mizoguchi, H., Tubbs, R. IX, Wiltrout, R. H., O’Shea, J. J., Kudoh, S., Klein, E., Bukowski, R. M. & Ochoa, A. C. (1993) Loss of T-cell receptor < chain and ~56”’ in T-cells infiltrating human renal cell carcinoma. Cancer Res. 53: 56135616. Rosenberg, S. A., Speiss, P., & Lefreniere, R. (1986) A new approach to the adoptive immunotherapy of cancer with tumor infiltrating lymphocytes. Science 233: 1318-l 321. Whiteside T. L. (1990) Human tumor-infiltrating lymphocytes and their characterization. In Lotzova, E. 81 Herberman, R. B. (eds) lnterleukin-2 and Killer Cells in Cancer. Boca Raton: CRC Press, pp. 147-l 69. Kradin, R. L., Boyle, L. A., Preffer, F. I,, Blumberg, R., Schneeberger, E. E. 81 Kurnick, J. T. (1987) Tumor-derived interleukin-2 dependent lymphocytes in adoptive immunotherapy of lung cancer. Cancer lmmunol. lmmunother. 24: 76-86. Kurnick, J. T., Kradin, R. L., Blumberg, R., Schneeberger, E. E. & Boyle, L. A. (1986) Functional characterization of T lymphocytes propagated from human lung carcinomas. Clin. lmmunol. lmmunopathol. 38: 367-377. Maleckar, J. R., Friddell, C. S., Sferruzza, A., Thurman, G. B., Lewko, W. M., West, W. H., Oldham, R. K., Dillman, R. 0.. Yanelli, J. R., Barth, N. M. & Maleckar, J. R. (1989) Activation and expansion of tumor-derived activated cells for therapeutic use. J. Natl. Cancer Inst. 81: 1655-l 661. Oldham, R. K., Dillman, R. O., Yanelli, J. R., Barth, N. M., Maleckar, J. R., Sferruzza, A., West, W. H., Friddell, C. S., Lewko, W. M. & Thurman, G. B. (1991) Continuous infusion interleukin-2 and tumor-derived activated cells as treatment of advanced solid tumors: A national biotherapy study group trial. Mol. Biother. 3: 68-77. Hanson, J. P., Kurtz, J., Rohloff, C., Kabler-Babbit, C.. Aleem J. & Rausch, C. (1993) Recombinant interleukin-2 with tumor-infiltrating lymphocyte (TIL) for metastatic renal cell carcinoma. Proc. Am. Sot. Clin. Oncol. 12: 1379. Alexander, J., Rayman, P., Edinger. M., Connelly, R., Tubbs, R., Bukowski, R. & Finke, J. H. (1990) TIL from renal-cell carcinoma: Restimulation with tumor influences proliferation and cytolytic activity. lnt. J. Cancer 45: 119-I 25. Kradin, R. L., Kurnick, J. T., Lazarus, D. S., Preffer. F. I., Dubinett, S. M. & Pinto, C. E. (1989) Tumour-infiltrating lymphocytes and interleukin-2 in the treatment of advanced cancer. Lancet l(8638): 577-81. Topalian, S. L., Solomon, D. & Avid, F. P. (1988) Immunotherapy of patients with advanced cancer using tumor-infiltrating lymphocytes and recombinant interleukin-2.: A pilot study. J. Clin. Oncol. 6: 839-848. Fisher, B., Packard, B. S., Read, E. J., Carrasquillo, J. A., Carter, C. S. & Topalian, S. L. (1989) Tumor localization of adoptively transferred Indium-111 labeled tumor infiltrating lymphocytes in patients with metastatic melanoma. J. Clin. Oncol. 7: 250-258. Mukherji, B., Arnbjarnarson, O., Spitznagle, L. A., Kalish, R. I., Hoffman, J., Ergin, M. T., Wilhelm, S. A., Nashed, A. L. & Guha, A. (1988) Imaging pattern of previously in vitro sensitized and interleukin-2 expanded autologous lymphocytes in human cancer. Nucl. Med. Biol. 15: 419-425. Topalian, S. L., Muul, L. M., Solomon, D. & Rosenberg, S. A. (1987) Expansion of human tumor infiltrating lymphocytes for use in immunotherapy trials. J. lmmunol. Meth. 102:127-38. Carpinito, G., Levine, S., Hamilton, D., Krane, R. J. & Osband, M. E. (1986) Successful adoptive immunotherapy of cancer using in vitro immunized antologous lymphocytes and cimetidine. Surg. Forum 27: 418421. Osband, M. E. & Krane, R. J. (1988) Autolymphocyte therapy: previous experience and future prospects. Pathol. lmmunopathol. Res. 7: 483-490. Ross, S., & Osband, M. E. (1989) Autolymphocyte therapy: an outpatient, low-toxicity approach to adoptive immunotherapy. Cancer Met. Rev. 8: 186-l 96. Krane, R. J., Carpinito, G. A., Ross, S. D., Lavin, P. T. & Osband, M. E. (1990) Treatment of metastatic renal cell carcinoma with antolymphocyte therapy: low toxicity outpatient approach to adoptive immunotherapy without use of in vitro interleukin-2. Urology 35: 417425. Gold, J. E. & Osband, M. E. (1994) Autolymphocyte therapy: I. In vivo tumor-specific adoptive cellular therapy of murine melanoma and carcinoma using ex vivo activated memory T-cells. J. lmmunother. in press.

ADOPTIVE 62.

63.

64.

65.

66.

67.

67a.

68.

69.

70.

71.

72.

73.

74.

75.

76.

CELLULAR

THERAPY

OF GU CANCER

187

Gold, J. E., Masters, T. R. & Osband, M. E. (1993) Autolymphocyte Therapy (ALT) of human renal cell carcinoma (RCC): Demonstration of anti-tumor cytotoxicitv (ATC) by ex viva activated memory T-cells and potentiation by cis-diamminedichloroplatinum(ll) (CDDP). Proc. Am. Sot. C/in. Oncol. 2: 740. Celis, E., Bolwerk, A., Clarke, J., Goodwin, J., Krane, R. J. & Osband, M. E. (1991) The immunologic mechanism of autolymphocyte therapy in the successful treatment of renal cell carcinoma (RCC) is the infusion of activated memory T-cells. J. Ural. 145: 339A. Crossland, K. D., Lee, V. K., Chen, W.. Riddell, S. R., Greenberg, P. D. & Cheever, M. A. (1991) T cells from tumor-immune mice nonspecifically expanded in v&o with anti-CD3 plus IL-2 retain specific function in vitro and can eradicate disseminated leukemia in vivo. J. lmmunol. 146: 44144422. Osband, M. E., Lavin, P. T., Babayan. R. K., Graham, S., Lamm, D. L., Parker, B., Sawczuck, I., Ross, S. & Krane, R. J. (1990) Effect of autolymphocyte therapy on survival and quality of life in patients with metastatic renal cell carcinoma. Lancet, 335: 994-999. Graham, S., Babayan, R. K., Lamm, D. L., Sawczuck, I., Ross, S. D., Lavin, P. T., Osband, M. E. & Krane, R. J. (1993) The use of ex viva-activated memory T-cells (autolymphocyte therapy) in the treatment of metastatic renal cell carcinoma: final results from a randomized, controlled, multisite study. Semin. Ural. 11: 27-36. Wang, J. C. L., Walle, A., Novogrodsky. A., Suthanthiran, M., Silver, R. T. & Bander, N. H. (1989) A phase II clinical trial of adoptive immunotherapy for advanced renal cell carcinoma using mitogenactivated antologous leukocytes and continuous infusion interleukin-2. J. C/in. Oncol. 7: 18851893. Ikarashi, H., Fujita, K., Takakuwa, K., Kodama, S., Tokunaga, A., Takahashi, T. &Tanaka, K. (1994) lmmunomodulation in patients with epithelial ovarian cancer after adoptive transfer of tumourinfiltrating lymphocytes. Cancer Res. 54: 190-I 96. Osband, M., Sewall, D. & Lavin, P. (1988) Autolymphocyte therapy (ALT) of renal cell carcinoma (RCC): clinical results and correlation of survival with serum interleukin-2 receptor (IL-2R) and immunosuppressive acidic protein (IAP). F’roc. Am. Sot. C/in. Oncol. 7: 491. Osband, M. E., Rosenthal, R. E. 81 Lavin, P. T. (1988) Autolymphocyte therapy (ALT) of renal cell carcinoma (RCC): correlation of survival with phenotype of the in vitro immunized lymphocytes. Proc. Am. Assoc. Cancer Res. 29: 1578. Crump, W. L. Ill, Owen-Schaub, L. B. & Grimm, E. A. (1989) Synergy of human recombinant interleukin-1 with interleukin-2 in the generation of lymphokine-activated killer cells, Cancer Res. 49: 149-I 57. Hemler, M. E., Jacobson, J. G., Brenner, M. B., Mann, D. & Strominger, J. L. (1985) VLA-1: a Tcell surface antigen which defines a novel late stage of human T-cell activation. Eur. J. Immunol. 15: 502-510. Ross, S., & Osband, M. (1989) Treatment of metastatic renal cell carcinoma (RCC) with autolymphocyte therapy: correlation between survival and the number of infused lymphocytes. FASEB 3: A825. Bianchi, R., Romani, L., Puccetti, P. & Fioretti, M. C. (1988) Induction of tumor suppression and delayed-type footpad reaction by transfer of lymphocytes sensitized to a xenogenized tumor variant. lnt. J. Cancer 42: 71-78. Puccetti, P., Bianchi, R., Romani, L., Cenci, E. & Fioretti, M. C. (1989) Delayed-type hypersensitivity to tumor antigens co-expressed with immunogenic determinants induced by xenogenization. Int. J. Cancer 43: 279-286. Gastl, G., Finstad, C. L., Guarini, A., Bosl, G., Gilboa, E. & Bander, N. H. (1992) Retroviral vector-mediated lymphokine gene transfer into human renal cancer cells. Cancer Res. 52: 62296236.

Itaya, T., Fearon, E., Fiesinger, T., Hunt, B., Vogelstein, B. & Frost, P. (1991) lmmunogenicity of a non-class I MHC expressing murine tumor transfected with the influenza virus hemagglutinin or murine interleukin-2 genes. Cancer Immunol. lmmunother. 33: 267-273. 77. Schoof, D. D., Jung, S. E. & Eberlein, T. J. (1989) Human tumor-infiltrating lymphocyte (TIL) cytotoxicity facilitated by anti-T-cell receptor antibody. Int. J. Cancer 44: 219-225. 77a. Townsend, S. E. & Allison, J. P. (1993) Tumor rejection after direct costimulation of CD8’ T cells by B7-transfected melanoma cells. Science 259: 368-370. 78. Finke, J. H., Rayman, P., Alexander, J., Edinger, M., Tubbs, R. R. & Bukowski, R. M. (1990)

188

79.

80.

81.

82.

83.

84.

85.

86. 87.

88. 89.

90.

91.

92.

93. 94. 95.

96.

97.

J. E. GOLD

ETAL.

Characterization of the cytolytic activity of CD4’ and CD8’ tumor-infiltrating lymphocytes in human renal cell carcinoma. Cancer Res. 50: 23632670. Schoof, D. D., Selleck, C. M., Massaro, A. F., Jung, S. E. & Eberlein, T. J. (1990) Activation of human tumor-infiltrating lymphocytes by monoclonal antibodies directed to the CD3 complex. Cancer Res. 50: 1138-l 145. Ochoa, A. C., Hasz, D. E., Rezonzew, R., Anderson, P. M. & Bach, F. M. (1989) Lymphokineactivated killer activity in long-term cultures with anti-CD3 plus interleukin 2: identification and isolation of effector subsets. Cancer Res. 49: 963-970. Bukowski, R. M., Sharfman, W., Murthy, S., Rayman, P., Tubbs, R., Alexander, J. & Finke, J. H. (1991) Clinical results and characterization of tumor-infiltrating lymphocytes with or without recombinant interleukin 2 in human metastatic renal cell carcinoma. Cancer Res. 51: 41994208. Yoshizawa, H., Chang, A. E. & Shu, S. (1991) Specific adoptive immunotherapy mediated by tumor-draining lymph node cells sequentially activated with anti-CD3 and IL-2. J. lmmunol. 147: 729-737. Chou, T., Chang, A. E. & Shu, S. (1988) Generation of therapeutic T lymphocytes from tumorbearing mice by in vi00 sensitization: culture requirements and characterization of immunologic specificity. J. lmmunol. 140: 2453-2461, Wong, J. T., Pinto, C. E., Gifford, J. D., Kurnick, J. T. & Kradin, R. L. (1989) Characterization of the CD4’ and CD8’ tumor infiltrating lymphocytes propagated with bispecific monoclonal antibodies. J. lmmunol. 143: 3404-3412. Perez, P., Hoffman, R. W., Titus, J. A. & Segal, D. M. (1986) Specific targeting of human blood T cells by heteroaggregates containing anti-T3 crosslinked to anti-target cell antibodies. J. Exp. Med. 163: 166-l 80. Liu, M. A., Nussbaum, S. R. & Eisen, N. H. (1988) Hormone conjugated with antibody to CD3 mediates cytotoxic T cell lysis of human melanoma cells. Science 239: 3G32. Fishleder, A. J., Finke, J. H., Tubbs, R. & Bukowski, R. M. (1990) Induction by interleukin-2 of oligoclonal expansion of cultured tumor-infiltrating lymphocytes. J. Nat/. Cancer inst. 82: 124131. Barth, R. J. Jr, Bock, S. N., Mule, J. J. & Rosenberg, S. A. (1990) Unique tumor-associated antigens identified by tumor-infiltrating lymphocytes. J. lmmunol. 144: 1531-I 539. Rosenberg, S. A., Aebersold, P., Cornetta, K., Kasid, K., Morgan, R. A., Moen, R., Morecki, S.. Karson, E.. Kasid, A., Blaese, R. M. & Anderson, W. F. (1990) Gene transfer into humans: Immunotherapy of patients with advanced melanoma using tumor-infiltrating lymphocytes modified by retroviral gene transduction. N. Engl. J. Med. 323: 570-575. Morecki S, Karson E, Cornetta K, Kasid A, Aebersold P, Blaese RM, Aebersold, P., Cornetta, K., Kasid, K., Morgan, R. A., Moen, R., Anderson, W. F. & Rosenberg, S. A. (1991) Retrovirusmediated gene transfer into CD4’ and CD8’ human T-cell subsets derived from tumor-infiltrating lymphocytes and peripheral blood mononuclear cells. Cancer Immunol. lmmunother. 32: 342-349. Fearon, E. R., Pardoll, D. M., Itaya, T., Golumbeck, P., Levitsky, H. I., Simons. J. W. & Vogelstein, B. (1990) Interleukin-2 production by tumor cells bypasses T helper function in the generation of anti-tumor responses. Ce// 60: 397408. Golumbeck, P. T., Lazenby, A. J., Levitsky, H. I., Jaffee, L. M., Karasuyama, H., Baker, M. & Pardoll, D. M. (1991) Treatment of established renal cell cancer by tumor cells engineered to secrete interleukin-4. Science 254: 713-716. North, R. J. (1984) y-irradiation facilitates the expression of adoptive immunity against established tumors by eliminating suppressor T cells. Cancer lmmunol. lmmunofher. 16: 175-l 83. North, R. J. (1982) Cyclophosphamide-facilitated adoptive immunotherapy of an established tumor depends on elimination of tumor-induced suppressor T cells. J. Exp. Med. 55: 1063-l 075. Klarnet, J. P., Kern, D. E., Okuno, K., Cheever, M. A. & Greenberg, P. D. (1990) Adoptive transfer of T cell for therapy of disseminated leukemia: Antigen specificity and function of tumor-reactive T cells. In Lotzova, E. & Herberman, R. B. (eds) lnterleukin-2 and Killer Cells in Cancer. Boca Raton: CRC Press pp. 200-222. Cameron, R. B., Speiss, P. J. & Rosenberg, S. A. (1990) Synergistic antitumor activity of tumorinfiltrating lymphocytes, interleukin-2, and local tumor irradiation. Studies on the mechanism of action. J. Exp. Med. 171: 249-260. Mitchell, M. S. (1989) Combining chemotherapy and biomodulation in the treatment of cancer. In

ADOPTIVE

98.

99.

100.

101.

102.

103.

104.

105. 106.

107. 108.

109.

110.

111.

112.

113.

114.

115.

CELLULAR

THERAPY

OF GU CANCER

189

Metzgar, R. S. & Mitchell, M. S. (eds) Human Tumor Antigens and Specific Tumor Therapy. New York: Alan R. Liss, pp. 345-355. Gold, J. E., Malamud, S. C., Osband, M. E., LaRosa, F. & Seder, R. (1993) Adoptive chemoimmunotherapy for the treatment of relapsed and refractory solid tumors using ex vivo activated memory T-cells (autolymphocyte therapy) and cyclophosphamide. J. Immunother. 13: 213221. Rosenberg, S. A., Packard, B. S., Aebersold, P. M., Solomon, D., Topalian. S. C., Toy, S. T., Simon, P., Lotze, M. T., Yang, J. C.. Seipp. C. A., Simpson, C., Carter, C.. Bock, S., Schwartzentruber, P., Wei, J. P. 81 White, D. E. (1988) Use of tumor-infiltrating lymphocytes and interleukin-2 in the immunotherapy of patients with metastatic melanoma. N. Engl. J. Med. 319: 1676-l 683. Formelli, F., Rossi, C., Sensi, M. L. & Parmiani, G. (1988) Potentiation of adoptive immunotherapy by cis-diamminedichloroplatinum (II), but not by doxorubicin, on a disseminated mouse lymphoma and its association with reduction of tumor burden. Int. ./. Cancer 42: 952-959. Mizutani, Y., Nio, Y. & Yoshida, 0. (1992) Modulation by cis-diamminedichloropaltinum(ll) of the susceptiblities of human T24 lined and freshly separated autologous urinary bladder transitional carcinoma cells to peripheral blood lymphocytes and lymphokine activated killer cells. J. Ural. 147: 505-513. Trasman. J., Higuchi, C. M., Thompson, J. A., Gillis, S., Lindgren, C. G. 81 Kern, D. E. (1990) Enhancement by interleukin-4 of interleukin-2 or antibody-induced proliferation of lymphocytes from interleukin-2 treated cancer patients. Cancer Res. 50: 1160-I 167. Chikkola, N. F., Lewis, I., Ulchaker, J., Stanky, J., Teibbs, R. & Finke, J. H. (1990) Interactive effects of a-interferon A/D and interleukin 2 on murine lymphokine-activated killer activity: Analysis at the effector and precursor level. Cancer Res. 50: 1176-l 184. Yang, S. C., Fry, K. D., Grimm, E. A. & Roth, J. (1990) Successful combination immunotherapy for the generation in vivo of antitumor activity with anti-CD3, interleukin 2. and tumor necrosis factor ct. Arch. Stag. 125: 220-225. Vander Woude, D. L., Wagner, P. D., Shu, S. & Chang, A. E. (1991) Enhanced antitumor reactivity of tumor-sensitized T cells by interferon alfa. Arch. Surg. 126: 307-314 Melder, R. J., Whiteside, T. L., Vujanovic, N. L., Hiserodt, J. C. & Herberman, R. B. (1988) A new approach to generating antitumor effecters for adoptive immunotherapy using human adherent lymphokine-activated killer cells, Cancer Res. 48: 3461-3470 Springer, G. F. (1984) T and Tn. general carcinoma autoantigens. Science 224: 1198-2004. Ghazizadeh, M., Kagawa, S., Izumi, K. 81 Kazuo, K. (1984) lmmunohistochemical localization of T antigen-like substance in benign hyperplasia and adenocarcinoma of the prostate. J. Ural. 132: 1127-l 135. Radzikowski, C. Z., Steuden, I., Wiedlochar, A., Kieler, J. & Tromholt, V. (1989) The ThomsenFriedenreich antigen on human urothelial cell lines detectable by peanut lectin and monoclonal antibody raised against human glycophorin A. Anticancer Res. 9: 103-l 06. Stamey, T. S., Yang, N., Jay, A. R., McNeal, J. E., Freiha, F. S. & Redwine, E. (1987) Prostatespecific antigen as a serum marker for adenocarcinoma of the prostate, N. Engl. J. Med. 317: 909916. Henningsson, C. M., Selvaraj. S., MacLean, G. D., Suresh, M. R., Noujaim, A. A. & Longenecker, B. M. (1987) T cell recognition of a tumor-associated glycoprotein and its synthetic carbohydrate epitopes: Stimlation of anticancer T cell immunity in vivo. Cancer lmmunol. Immunother. 25: 231238. Fung, P. Y. S., Madej, M., Koganti, R., & Longenecker, B. M. (1990) Active specific immunotherapy of a murine mammary adenocarcinoma using synthetic tumor associated glycoconjugate. Cancer Res. 50: 43084316. Osband, M. E., Plummer, J. M., Goodwin, J. J., Hamilton, D. & Abulafia, R (1990). In vitro production of T-cells specifically immunoreactive against prostate specific antigen: and approach to the antigen-specific adoptive immunotherapy of prostate cancer. Proc. Am. Assoc. Cancer Res. 31: 278. Sznol, M., Clark, J. W., Smith, J. W., Steis, R. G., Urba, W. J. & Rubinstein, L. V. (1992). Pilot study of interleukin-2 and lymphokine-activated killer cells combined with immunomodulatory doses of chemotherapy and sequenced with interferon alfa-2a in patients with metastatic melanoma and renal cell carcinoma. J. Nat/. Cancer inst. 84: 929-938. Hanson, J., Petit, R., Walker, M., Kurtz, J., Rohloff, C. & Kabler-Babbitt, C. (1992) Tumor infiltrating

190

116.

117.

118.

119.

120.

121.

122.

J. E. GOLD

ETAL.

lymphocyte (TIL) therapy for metastatic renal cancer (RC) using interleukin-2 (IL-2). Proc. Am. Sot. Clin. Oncol. 11: 682. Belldegrun. A., Pierce, W., deKernion, J. & Figlin, Ft. (1993) The treatment of metastatic renal cell carcinoma with tumor infiltrating lymphocytes (TIL) and interleukin-2 containing regimens: The UCLA experience. J. Urd. 80: 352A (abstract). Rosenberg, S. A., Schwarz, S. L. & Spiess, P. J. (1988) Combination immunotherapy for cancer: Synergistic antitumor interactions of interleukin-2, alfa interferon, and tumor-infiltrating lymphocytes. J. Natl. Cancer Inst. 80: 1393-I 399. Peyret, C., Bochner, B. H., Lee, T. Y., Koo, A. S., deKernion, J. B. & Belldegrun A. (1992) Regulatory effects of interleukin-4 on tumor-infiltrating lymphocytes derived from human renal cell carcinoma. J. Surg. Res. 53: 602-608. Gelber, C., Eisenbach, L., Feldman, M., & Goodenow, R. S. (1992) T-cell subset analysis of Lewis lung carcinoma tumor rejection: Heterogeneity of effecters and evidence for negative regulatory lymphocytes correlating with metastasis. Cancer Res. 50: 6507-6514. Wang, P., Vanky, F., Li, S. L., Vegh, Z., Persson, U. 81 Klein, E. (1991) Expression of MHC-class-l antigens in human carcinomas and sarcomas analyzed by isoelectric focusing. ht. J. Cancer 6: 106-l 12. Zhang, Z. L., Kurtzberg, K., Osband, M., Miesowicz, F. & B&bit, B. (1993) Synergistic effects of autologous cytokines and OKT3 in the ex vivo activation of memory T lymphocytes utilized in adoptive immunotherapy of metastatic renal cell carcinoma. J. lmmunol. 150: 1247. Chang, A. E., Yoshizawa, H., Sakai, K., Cameron, M. J., Sondak, V. K. & Shu, S. (1993) Clinical observations on adoptive immunotherapy with vaccine-primed T-lymphocytes secondarily sensitized to tumor in vitro. Cancer Res. 53: 1043-I 050.