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β chain-CD3 protein complex defect in systemic lupus erythematosus: T-cell function
T-Cell Receptor a/P Chain-CD3 Protein Complex Defect in Systemic Lupus Erythematosus: T-Cell Function SYEDRAZIUDDIN,Ph.D., MANSOURAHMEDAL-JANADI,M.D., ABDULHAMIDA. ALWABEL,M.D., Abha. SaudiArabia
We describe the first case of systemic lupus erythematosus (SLE) in which peripheral blood T cells were deficient in cell surface expression of T-cell receptor (Y/O chain (Tcm) and the CD3 protein. Because of the uncommon phenotype and because of the notion that coexpression of Tcw and CD3 is essential for antigen-specific T-cell function, in vitro functional assays were performed, showing a highly decreased proliferative responee to anti-CD3 antibody and other T-cell mitogens, deficient interleuhin-2 (IL-2) secretion, and impaired function to respond in autologous and allogeneic mixed lymphocyte reactions. However, the helper-inducer function of T cells was unaffected by deficient expression of the T&@/CD3 protein complex. The relative iucrease of CD4+ CDw23+ helperinducer subsets in T cells accounted for elevated secretion of two terminal B-cell stimulating factors, B-cell growth factor (BCGF) and B-cell differentiation factor (BCDF). Hence, our results suggest that the regulation of secretion of lymphohines, I&2, and BCGF and BCDF is independently controlled in T cells, and this case ihstrates the pathologic sequelae of a unique defect in T celh~ characteristic of SLE.
From the Department of Clinical Immunology (SR). and the Department of Internal Medicine (MAAl, AAA). Rheumatology Section, King Saud University, College of Medicine, Abha, and Asir Central Hospital, Abha, Saudi Arabia. Requests for reprints should be addressed to Syed Raziuddin. Ph.D., Department of Clinical Immunology, College of Medicine, Abha. P.O. Box 641, Abha, Saudi Arabia. Manuscript submitted August 28, 1990. and accepted in revised form April 15, 1991.
ystemic lupus erythematosus (SLE) is a multisystem autoimmune disease characterized by autoantibody production, hypergammaglobulinemia, defective interleukin-2 (IL-2) secretion, Bcell hyperactivity, and T-cell abnormalities [l-8]. Genetic and environmental factors contribute to these abnormalities of immune function. T lymphocytes have been implicated in an array of immunoregulatory phenomena in normal and abnormal immune systems. The regulation of immune response is largely a reflection of cumulative interactions among T cells, each with its own function and distinguishable from other T cells due to differences in surface antigen expression. Mature CD3+ T cells express either CD4+ or CDS+ cell surface glycoproteins. The protein products of the T-cell receptor (TcR) (Yand /3 genes and of y and 6 genes associate, forming a TcRc@ or TcR$ heterodimer [g-13]. TcRa/3 protein chains are linked by a disulfide bridge and are present on more than 90% of the peripheral blood CD3+ T cells [g-11]. A minor subpopulation of circulating T cell8 bear an alternative, TcRr6, but most of these cells express neither CD4 nor CD8 [9,13]. A functional division of labor exists between the TcR& and the CD3 protein, and coexpression of these components is essential for antigen-specific T-cell function [10,13,14]. T cells cannot recognize an antigen unless the TcRa&CD3 complex is correctly assembled and transported to the plasma membrane8 [13,14]. Quantifying TcR@/CDS+ T cells has been important in the characterization of immunologic abnormalities in patients with T-cell defects. Recently, congenital and acquired immunodeficiency diseases have been reported with either discordant or deficient expressions of TcR& and CD3 proteins on T cells [15-201, but to our knowledge, a defect in the TcRcrSICD3 protein on T cells of patients with SLE has not been reported previously. In this article, we describe a patient with SLE who had a defect in peripheral blood T cells in expression of TcRct$ and CD3 protein. For comparisons with these TcR&/CD3-negative T cells, we have also included a patient with typical SLE as the patient control. Functional characteristics of these T cells are presented here.
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PATIENTS AND METHODS Patient The patient was a 36-year-old nonworking Indian female expatriate living in Saudi Arabia since 1978. She was the wife of a physician working at one of the community hospitals in Abha. In November 1988, she was referred to our hospital for evaluation of high fever and polyarthritis. At that time she had a positive antinuclear antibody (ANA) titer of 1:360, but anti-nDNA was negative. Serum complement levels were slightly low-C3, 51 mg/dL (normal range: 60 to 170 mg/dL) and C4,17 mg/dL (normal range: 19 to 75 mg/dL)-but serum immunoglobulins were elevated: IgG 1,470 mg/dL (normal 780 to 1,200 mg/dL), IgA 285 mg/dL (normal 80 to 190 mg/dL), and IgM 260 mg/dL (normal 70 to 180 mg/dL). Laboratory studies revealed a normal hemoglobin level, liver enzyme values, white blood cell count, and platelet count but an elevated erythrocyte sedimentation rate (ESR) of 65 mm/h. At this time, T-cell studies were not done. She was treated with antirheumatic drugs with some improvement. Since then the patient has occasionally complained of fever, headache, and arthritis. In September 1989, she suddenly presented with a clear butterfly rash on her face, fever, headache, polyarthritis, mild alopecia, and photosensitivity. At this time she had a high anti-nDNA titer at 1:2,560, ANA titer of 1:1,280, and hypocomplementemia (C3,19 mg/dL; C4,6 mg/dL). Her ESR was 110 mm/h, hemoglobin was 9.2 mg/dL, and white blood cell count was 2.5 X log/L with 24% lymphocytes, 61% segmented neutrophils, 11% band forms, and 4% monocytes. Serum creatinine level, liver enzyme values, and results of urinalysis were normal. There was no serologic evidence of human immunodeficiency virus, hepatitis, Epstein-Barr virus, or cytomegalovirus infection. Liver, spleen, and lymph nodes were not enlarged. There was no evidence of malignancy. She was diagnosed as having SLE as per the American Rheumatism Association’s (ARA) revised criteria [21]. At this time, detailed immunologic studies were performed on blood samples. She was given 80 mgld of prednisolone. Over the next 10 days, her condition deteriorated and she developed nephrotic-range proteinuria followed by thrombocytopenia (platelet count 105 X log/L) and memory disturbances. A computed tomographic brain scan showed multiple infarcts. She died suddenly in the intensive care unit 1 week later from massive brain infarction and septic shock. Patient Control We examined a 29-year-old female Saudi patient with SLE who fulfiied the ARA revised criteria
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[21]. This patient had positive ANA (1:640 titer) and anti-nDNA (1:1,280 titer), decreased complement levels (C3,27 mg/dL; C4,12 mg/dL), and elevated serum IgG, IgA, and IgM levels. The clinical signs included a butterfly rash on the face, mild alopecia, polyarthritis, and thrombocytopenia. T-Cell Studies Peripheral blood mononuclear cells (MNC) were obtained by Histopaque-1077 (Sigma Chemical Co., St. Louis, MO) density gradient centrifugation. B and T cells were separated by the neuraminidasetreated sheep erythrocyte rosetting (E-rosette) technique [22]. CD4+ T cells were purified from Erosetting T cells using OKT8 monoclonal antibody solutions by a panning technique described earlier WI.
Lymphocyte Surface Marker Analysis Surface membrane phenotyping of MNC was determined with the OK and Leu series of monoclonal antibodies (Ortho Diagnostics, Raritan, NJ, and Be&on Dickinson, Mountain View, CA) by the standard indirect immunofluorescence technique [8,22]. The CD nomenclature and specificity of the monoclonal antibodies are shown in Table I. Proliferation and IL-2 Production The proliferative response to anti-CD3 monoclonal antibody (OKT3, 0.2 rg/mL), phytohemagglutinin (PHA, 2 rg/mL), and concanavalin A (Con A, 5 rg/mL) was determined on patient and control T cells as described earlier [7,22]. In brief, 2.5 X lo5 purified E-rosetting T cells were cultured with antiCD3, PHA, or Con A for 72 hours at 37OC in RPMI1640 medium, and incorporation of 3H-thymidine (3H-TdR) into DNA was determined. IL-2 activity defined as U/mL from the supernatants of T cells (2.5 X 105) cultured for 48 hours with mitogens used in the proliferative response was measured in a standard proliferation assay using an IL-2-dependent murine cytotoxic T-lymphocyte cell line as described [22]. B-Cell Growth Factor (BCGF) and B-Cell Differentiation Factor (BCDF) Activity T cells (5 X 105) were incubated in 1 mL of RPMI1640 medium with 1% fetal calf serum, 5 X 10s5 2mercaptoethanol, and 2 rg/mL of PHA. After 72 hours, the supernatants were recovered and were analyzed for BCGF and BCDF activity. BCGF activity from the T-cell supernatants was determined with normal B cells by incorporation of 3H-TdR according to the standard co-stimulator assay [22]. BCDF activity from the T-cell supernatants was
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determined by using Staphylococcus aureus Cowan I (SAC&stimulated normal B cells. B cells were cultured with 0.005% SAC and T-cell supernatanta for 7 days, and were then assayed for IgM and IgG secretion by enzyme-linked immunosorbent assay as shown [22]. Autologous Mixed Lymphocyte Reaction (AMLR) and Allogeneic Mixed Lymphocyte Reaction (MLR) T (92% to 95% E-rosetting and CD2+ cells) and non-T cells (62% to 74% CD24+ B cells, 15% to 18% CDll+ monocytes, and 1% to 4% CD2+ cells) were isolated from MNC by the E-rosetting technique [22]. The non-T cells were pretreated with 50 rg/mL of mitomycin-C (Sigma) for 45 minutes at 37OC. The AMLR and MLR activity of T cells was determined by culturing equal numbers of T cells with mitomycin-C-treated autologous non-T cells (for AMLR) or mitomycin-C-treated allogeneic normal non-T cells (for MLR) for 7 days at 37OC in a proliferation assay [7,22].
RESULTS The surface phenotypes of MNC from two SLE patients, one with deficient (patient) and the other with normal (patient control) TcRc@ and CD3 protein on T cells, were analyzed (Table I). This patient’s T cells lacked expression of both TcRaj3 and CD3 antigens as determined by staining with TcRla/3 (WT31) and OKT3/Leu-4 monoclonal antibodies. This observation of the deficiency of TcRa/3 and CD3 antigen from the patient was confirmed on several occasions at a l-day interval on freshly isolated MNC and E-rosetting purified T cells by using a range of TcR-l@ (WT31), OKT3, and Leu-4 monoclonal antibody concentrations that brightly labeled T cells from the patient control and normal controls. However, both the patient and the patient control had a norhal percentage of the CD2+ and E-resetting+ pan-T cells. They had increased CD4+ and decreased CD8+ T cells. Most patients with SLE we have investigated earlier [7] had an increased percentage of HLA-DR+ (Ia) cells in MNC, which was due to expression of these antigens on T cells. Of interest, HLA-DR+ cells were normal in MNC of our TcR@/CD3-negative patient with SLE, which probably accounts for expression of these antigens on B cells and some monocytes. However, as expected, an increased percentage of HLA-DR+ cells was present in MNC of our patient control. CD4+ CD45R+ (suppressorinducer) cells were decreased with a concomitant increase in CD4+ CDw29+ (helper-inducer) cells in the patient and patient control as compared with normal controls. Further examination of the puri-
TABLE I Phenotypic Characterization of Peripheral Blood Mononuclear Cells % mAbSpecificity Patient Normal Patient Control Control* T-cell receptor(TcR-l&WT31)
0
71
68 + 6
CD1 (OKT6,thymocytes)
2
4
622
CD2 (OKTll,panT)
74
73
7124
CD3cOKT3,panT)
1
70
72 2 4
CD3 (Leu-4,panT)
0
73
73 + 5
CD4 (OKT4, helperT)
61
54
45 + 4
CD8 (OKT8,suppressorT)
12
17
262
CD24(OKB2,Bcells)
11
13
1422
6
5
922
11
43
1223
E-rosettingt (pan-T-cellantigen)
73
72
72 + 4
CD4tCD45Rtt(OKT4,2H4, suppressor-inducer)
16
21
46 k 6
CD4tCDw29tt(OKT4,4B4, helper-inducer)
78
69
41k4
CDll(OKMI,monocytes) HLA-DR (OKlal,
la antigens)
3
4b = moncxlonal antlbody. lormal age-matched controls (n = 4) ? SD. ., neep erytnrocyte rosettmg cells. tmfied CD4t T cells were stained with monoclonal antibodies to CD45R L?H4) and CDw29 84).
fied CD4+ T-cell subsets from the patient with monoclonai antibodies to TcR@ and CD3 protein confirmed negative staining of these subsets for the TcRc@/CD3 protein complex. The function of purified E-rosetting T cells of the patient, patient control, and normal controls in proliferative response and IL-2 secretion is shown in Table II. As shown, the TcR@/CD3-deficient patient’s T cells demonstrated a highly decreased proliferative response to anti-CD3 monoclonal antibody, and a decreased response to PHA and Con A. Similarly, the culture supernatanta of this patient’s T cells produced little IL-2 in response to stimulation with these mitogens. These functional characteristics of T cells from the patient control were higher than in the patient but lower than in normal controls. Upon stimulation with antigens, normal T cells produce various lymphokines such as IL-2, BCGF, and BCDF. The BCGF and BCDF are of crucial importance in humoral immune response and antibody synthesis, which provides a stimulus for differentiation of B cells into antibody-secreting plasma cells. Of interest, although IL-2 activity in the TcRa/3/CD3-deficient patient’s T cells was highly
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TABLE II Functional Characteristicsof T Cells in Proliferative Responseand Interleukin-2 (IL-21 Secretion T Cells
Anti-CD3
PHA
Con A
1.8 + 0.6 6
16.6 + 2.2 21
9.2 k 1.8
38.4 + 4.3 37
55.6 f 4.0 46
32.6 ? 3.1 35
112.4 + 12.3
174.6 ? 17.8
70
93
Patient Proliferation* IL-2 activity+ Patient control Proliferation IL-2 activity Normal control Proliferation IL-2 activity
17
152.6
? 14.5
78
Phfl = pnyronemagglunnln; Mn A = concanavalln A. *Prokferabon (cpm x lo? was determined by culturing T cells with mitogens for 72 hours and incorporatIon of 3H-thymldme, cpm + SD of triplicate cultures after subtracting the background counts. 111-2activity (U/mL) was determined from the supernatants of T cells cultured for 48 hours with mitogens in a proliferationassay usingthe IL-2-dependent cytotoxic T-lymphocyte cell line.
decreased, such cells demonstrated increased BCGF and BCDF activity as compared with that in the patient control’s and normal control’s T cells (Table III). Because of functional and lymphokine abnormalities present in the TcRo$/CD3-deficient patient’s T cells and because of the notion that AMLR and MLR reflect an immunoregulatory mechanism that is important in SLE and other T-cell-mediated diseases, we have also examined the capacity of this patient’s T cells to proliferate in AMLR and MLR. AB shown in Table IV, highly deficient AMLR and MLR activity of the TcR&CD3-deficient patient’s T cells was evident as compared with that in the patient control and normal controls.
COMMENTS We describe a patient with SLE whose peripheral blood T cells expressed CD2+ and E-rosetting panT cells but lacked the normal TcRapKD3 surface protein complex. These TcRa$/CD3 protein complex-negative T cells expressed either the CD4+ or CD8+ phenotype. Expression of both TcRaS and CD3 proteins together on CD4+ or CD8+ subsets is
essential for antigen-specific T-cell function. As a result of deficient cell surface expression of the TcR@3/CD3 complex, T-cell proliferation in response to anti-CD3 monoclonal antibody, PHA, and Con A was impaired, as was IL-2 secretion. Likewise, these T cells showed a highly reduced capacity to proliferate when cultured with either autologous or allogeneic non-T cells in the standard AMLR and MLR assays. However, they were able to secrete elevated levels of two terminal B-cell stimulating factors, BCGF and BCDF. These data suggest that the regulation of IL-2 secretion and BCGF and BCDF secretion by T cells in SLE is independent. On the basis of these findings, we demonstrated that a loss of the TcR@/CD3 protein complex from T cells does not change their ability to influence B-cell function in antibody production, which is a pathologic sequela of T cells in SLE. Investigations of immune function in patients with SLE have revealed IL-2 deficiency and T-cell subset abnormalities [l-8]. In particular, those patients with renal disease and central nervous system involvement have an absolute depression of the CD4+ CD45R+ suppressor-inducer subsets in the circulation [8,23]. The essential findings in both of our SLE patients, the TcRa/.VCD3 protein complex-deficient patient and the patient control, have been depletion of CD4+ CD45R+ cells with an expansion of CD4+ CDw29+ cells. Thus these results parallel those reported earlier by us and others [8,23]. The precise mechanism of the decreased CD4+ CD45R+ cells in SLE is not very clear. However, sera of the patients with a loss of CD4+ CD45R+ cells could have anti-T-cell antibodies directed against this subset, and therefore this may result in their elimination from the blood [4]. Alternatively, the CD4+ CD45R+ cells may represent virgin or naive cells and the CD4+ CDw29+ cells memory [24], and the abnormality observed in SLE may represent a block in differentiation. It is likely that environmental factors such as viruses, ultraviolet light, drugs, or other insults could possibly contribute continuously to these T-cell subset abnor-
TABLE Ill B-Cell Growth Factor (BCGF)and B-Cell Differentiation Factor (BCDF)Activity of T Cells T Cells Stimulated With
Patient BCGF* (cpml
None
1,830
PHA
17,840
BCDF+ Ifi
IN
620 10,200
890 19,500
BCGF (cpm) 1,320 14,290
PatientControl BCDF IgG kM 530 8,760
BCGF (cpm)
NormalControl BCDF M kM
810
1,040
310
15,220
8,300
4,600
iA = phytohemagglutinin. ICGF activity, %thymidine incorporation, meanof triplicate cultures was examinedin a proliferationassay of B cells with T-cell-stimulated culture supernatants Data shown are in cpm. CDFactivity, I& and IgM secretion (ng/mL), was examined by culturing B cells with mitogen-stimulatedT-cell supernatants.
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T CELLS IN SLE / RAZIUDDIN ET AL
malities in the genetically predisposed host. The pathologic significance of this finding may be related directly to enhanced T-cell helper-inducer function by the CD4+ CDw29+ subsets on B-cell hyperactivity in autoantibody secretion, characteristic of this disease. Therefore, the increased secretion of BCGF and BCDF in the T cells of our TcRa/3/CD3 protein complex-negative patient should account for the increased representations of the CD4+ CDw29+ helper-inducer subsets in T-cell populations. The AMLR represents a proliferative response of T cells to self-Ia antigens and that class II major histocompatibility complex gene products alone on autologous non-T cells are capable of triggering this response [6,25]. Moreover, it would appear that the AMLR and MLR in humans are regulated by the same set of rules that govern antigen-specific T-cell activation. We show that the T cells of our TcRc&CD3 protein complex-negative patient did not proliferate either in AMLR or in MLR assays. However, the TcRc@/CD3-positive SLE patient’s T cells demonstrated lower AMLR and MLR activity than in normal controls, but produced higher activity than the TcRaglCD3negative cells. Thus, the variations seen between TcRc@/CD3-negative and positive T cells from the SLE patients suggest that these differences in the functional behavior may be due to the expression of TcR@/CD3 complexes. Genetic predisposition is known to play a crucial role in the development of SLE [26,27]; genetic factors may also contribute to some of the specific abnormalities in T cells. Family studies in the patient’s two healthy children (8-year-old girl and 6year-old boy) and a healthy sister (23 years old) demonstrated that these individuals had a normal percentage of TcR@/CDS-positive T cells in blood (data not shown). There is no family history of immunodeficiency or SLE. Therefore, it is most likely that deficiency of the TcRa@/CD3 protein complex on the T cells of this patient is an acquired abnormality that occurred during the development of SLE. Alternatively, the serum of this patient may have anti-TcRaj3 and anti-CD3 antibodies, which might block or modulate expression of the TcRatflICD3 protein complex on T cells. There are four groups of rearranging genes in human T cells that encode the T-cell receptors for antigens (TcRa, /3, y, and 6). T cells recognize antigens in the context of the major histocompatibility complex through CDS-associated receptors: (~0 chain on mature cells and y6 chain on immature cells [13,14]. Several lines of evidence indicate that expression of both TcR@ and CD3 protein complexes is essential for CD4+ helper/inducer and
TABLE IV Autologous Mixed LymphocyteReaction (AMLR) and Allogeneic Mixed LymphocyteReaction (MLR) Activity of T Cells 1 Cells From Subjects
3H-Thymidine AMLR*
Patient Patlent control Normal control
2.3 + 0.8 28.6 e 3.4 46.5 2 5.2
Incorporation
kpm x 103) MLR+ 1.7 2 0.5 23.2 2 2.8 39.5 2 3.7
I
*AMLR acbvlty was examined by culturing T cells with autologous mitomycln C-treated non-T cells. ‘MLR activity was examined by culturingT cells with mitomycin C-treated allogenelcnormal non-T cells. tData shown are mean r SD of triplicate cultures after subtracting the background counts (T cells cultured alone in medium).
CD8+ suppressor/cytotoxic T-cell function in an immune response [ll-141. Earlier, TcRc@/CD3 protein complex deficiency on T cells was reported from T-lineage lymphomas [16] and in a patient with severe combined immunodeficiency [17]. However, the discordant expression of TcRa@ and CD3 (TcRafl+/CD3-, TcRc+/CD3+) on T cells is a common finding in patients with juvenile rheumatoid arthritis [12], leprosy [ 151, non-Hodgkin’s lymphomas [ 161, Hodgkin’s disease [ 191, and ataxia telangiectasia [20], and has been reported in a patient with combined immunodeficiency with features of graft-versus-host disease [18]. In juvenile rheumatoid arthritis, TcRo$/CD3+ cells represent a notable fraction of spontaneously activated T cells at the site of the lesion in joints, which has led to the speculation that they are involved in the pathogenetic mechanisms occurring in this disease [12]. In conclusion, we have described a new population of human T cells, all with the TcR&CD3phenotype in the circulation of a patient with SLE. On the basis of the monoclonal antibody reactivity of MNC, these TcRo$VCD3- T cells may consist of two distinct subsets, a major population with the TcRaB-, CD3-, CD4+ phenotype and a minor population with the TcRa/3-, CD3-, CD8+ phenotype. The CD4+ populations may further be subdivided into TcR&-, CD3-, CD4+ CD45R+ and TcRafl-, CD3-, CD4+ CDw29+ subsets. Because of our patient’s death, we have not performed studies on T-cell receptor gene rearrangements, and have not determined whether these T cells express an alternative TcR$ chain instead of TcRc@. However, it is unlikely that this TcRaB/CD3T-cell population may express the TcRys chain. The existing evidence that TcRy&positive T cells express neither CD4 nor CD8 antigens lends support to this interpretation. On the basis of functional characteristics observed in proliferative response, AMLR and MLR assays, and lymphokine secretion, it was demonstrated that some usual functions of T cells
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in disease states are severely modified, whereas others are not. We believe that this may be due to the deficiency of TcRcq3 and CD3 on the surface of such cells. Whether such TcR&CD3protein complex-deficient T cells can be identified from immunodeficient cases and additional patients with SLE and other human autoimmune diseases in the future is awaited. In this regard, it will be of interest to monitor changes in the phenotype and pattern of response during the course of the disease. As more is learned about immunoregulatory circuits affecting T-cell receptor gene products in humans, the processes regulating these events in Tcell differentiation pathways may be better understood.
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10. Lanier LL, Federspiel NA, Ruitenberg JJ. The T cell antigen receptor complex expressed on normal peripheral blood CD4-, CD&T lymphocytes: a CD3 associated disulfide linked y chain heterodimer. J Exp Med 1987; 165: 1075-94. 11. Allison JP, Lanier LL. The T cell antigen receptor gamma gene: rearrangement and cell lineage. lmmunol Today 1987; 8: 293-313. 12. DeMaria A, Malnati M. Moretta A, et al. CD3+ 4- B- WT31- (T cell receptor y+) cells and other unusual phenotypes are frequently detected among spontaneously interleukin-2 responsive T lymphocyte present in the joint fluid in juvenile rheumatoid arthritis: a clonal analysis. Eur J lmmunol 1987; 17: 1815-g. 13. Allison JP. Lanier LL. Structure, function and serology of the T cell antigen receptor. Annu Rev lmmunol 1987; 5: 503-40. 14. Strominger JL. Developmental biology of T cell receptors. Science 198% 244: 943-50. 15. Modlin RL. Brenner MB, Krangel MS, Duby AD, Bloom BR. T cell receptors of human suppressor cells. Nature 1987; 329: 541-4. 16. Picker LJ, Brenner MB, Weiss LM, Smith SD, Warnke RA. Discordant expression of CD3 and T cell receptor beta chain antigens in T-lineage lymphomas. Am J Pathol 1987; 129: 434-40. 17. Alarcon B. Requeiro JR, Arnaiz-villena A, Terhorst C. Familial defect in the T cell receptor-CD3 complex. N Engl J Med 1988; 319: 1203-B. 16. Wirt DP. Brooks ED, Vaidys S. et a/. Novel T lymphocyte population in combined immunodeficiency with features of graft-versus host disease. N Engl J Med 1989; 321: 370-4. 19. Dallenbach FE, Stein H. Expression of T cell receptor j3 chain in Reed Sternberg cells. Lancet 1989; 2: 828-30. 20. Carboneri M, Cherchi M, Panganelli P, et al. Relative increase of T cells expressing the gamma/delta rather than the alpha/beta receptors in ataxia telangiectasia. N Engl J Med 1990; 322: 73-6. 21. Tan EM, Cohen AS, Fries JF, eta/. The 1982 revised criteria for the classification of systemic lupus erythematosus. Arthritis Rheum 1982; 25: 1271-7. 22. Raziuddin S, Teklu B. Severe T lymphocyte immunodeficiency associated with hypogammaglobulinemia: defective lymphokine secretion but enhanced autologous mixed lymphocyte reaction. J Clin lmmunol 1989; 9: 448-53. 23. Morimoto C. Steinberg AD, Letvin NL. et a/. A defect of immunoregulatory T cell subsets in systemic lupus erythematosus patients demonstrated with anti2H4 antibody. J Clin Invest 1987; 79: 762-B. 24.Sanders ME, Makgoba MW, Shaw S. Human naive and memory T cells: reinterpretation of helper-inducer and suppressor-inducer subsets. lmmunol Today 1988; 9: 195-9. 25. Weksler ME, Moody CE Jr, Kozak RW. The autologous mixed lymphocyte reaction. Adv lmmunol 1981; 31: 271-312. 26. Block SR. Lokshin MD, Winfield JB. eta/. Immunologic observations on 9 sets of twins either concordant or discordant for systemic lupus erythematosus. Arthritis Rheum 1976; 19: 545-54. 27. Sakane T. Murakawa Y, Suzuki N, et al. Familial occurrence of impaired interleukin-2 activity and increased peripheral blood B cells actively secreting immunoglobulins in systemic lupus erythematosus. Am J Med 1989; 86: 385-90.
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We describe the first case of systemic lupus erythematosus (SLE) in which peripheral blood T cells were deficient in cell surface expression of T-cell receptor (Y/O chain (Tcm) and the CD3 protein. Because of the uncommon phenotype and because of the notion that coexpression of Tcw and CD3 is essential for antigen-specific T-cell function, in vitro functional assays were performed, showing a highly decreased proliferative responee to anti-CD3 antibody and other T-cell mitogens, deficient interleuhin-2 (IL-2) secretion, and impaired function to respond in autologous and allogeneic mixed lymphocyte reactions. However, the helper-inducer function of T cells was unaffected by deficient expression of the T&@/CD3 protein complex. The relative iucrease of CD4+ CDw23+ helperinducer subsets in T cells accounted for elevated secretion of two terminal B-cell stimulating factors, B-cell growth factor (BCGF) and B-cell differentiation factor (BCDF). Hence, our results suggest that the regulation of secretion of lymphohines, I&2, and BCGF and BCDF is independently controlled in T cells, and this case ihstrates the pathologic sequelae of a unique defect in T celh~ characteristic of SLE.
From the Department of Clinical Immunology (SR). and the Department of Internal Medicine (MAAl, AAA). Rheumatology Section, King Saud University, College of Medicine, Abha, and Asir Central Hospital, Abha, Saudi Arabia. Requests for reprints should be addressed to Syed Raziuddin. Ph.D., Department of Clinical Immunology, College of Medicine, Abha. P.O. Box 641, Abha, Saudi Arabia. Manuscript submitted August 28, 1990. and accepted in revised form April 15, 1991.
ystemic lupus erythematosus (SLE) is a multisystem autoimmune disease characterized by autoantibody production, hypergammaglobulinemia, defective interleukin-2 (IL-2) secretion, Bcell hyperactivity, and T-cell abnormalities [l-8]. Genetic and environmental factors contribute to these abnormalities of immune function. T lymphocytes have been implicated in an array of immunoregulatory phenomena in normal and abnormal immune systems. The regulation of immune response is largely a reflection of cumulative interactions among T cells, each with its own function and distinguishable from other T cells due to differences in surface antigen expression. Mature CD3+ T cells express either CD4+ or CDS+ cell surface glycoproteins. The protein products of the T-cell receptor (TcR) (Yand /3 genes and of y and 6 genes associate, forming a TcRc@ or TcR$ heterodimer [g-13]. TcRa/3 protein chains are linked by a disulfide bridge and are present on more than 90% of the peripheral blood CD3+ T cells [g-11]. A minor subpopulation of circulating T cell8 bear an alternative, TcRr6, but most of these cells express neither CD4 nor CD8 [9,13]. A functional division of labor exists between the TcR& and the CD3 protein, and coexpression of these components is essential for antigen-specific T-cell function [10,13,14]. T cells cannot recognize an antigen unless the TcRa&CD3 complex is correctly assembled and transported to the plasma membrane8 [13,14]. Quantifying TcR@/CDS+ T cells has been important in the characterization of immunologic abnormalities in patients with T-cell defects. Recently, congenital and acquired immunodeficiency diseases have been reported with either discordant or deficient expressions of TcR& and CD3 proteins on T cells [15-201, but to our knowledge, a defect in the TcRcrSICD3 protein on T cells of patients with SLE has not been reported previously. In this article, we describe a patient with SLE who had a defect in peripheral blood T cells in expression of TcRct$ and CD3 protein. For comparisons with these TcR&/CD3-negative T cells, we have also included a patient with typical SLE as the patient control. Functional characteristics of these T cells are presented here.
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PATIENTS AND METHODS Patient The patient was a 36-year-old nonworking Indian female expatriate living in Saudi Arabia since 1978. She was the wife of a physician working at one of the community hospitals in Abha. In November 1988, she was referred to our hospital for evaluation of high fever and polyarthritis. At that time she had a positive antinuclear antibody (ANA) titer of 1:360, but anti-nDNA was negative. Serum complement levels were slightly low-C3, 51 mg/dL (normal range: 60 to 170 mg/dL) and C4,17 mg/dL (normal range: 19 to 75 mg/dL)-but serum immunoglobulins were elevated: IgG 1,470 mg/dL (normal 780 to 1,200 mg/dL), IgA 285 mg/dL (normal 80 to 190 mg/dL), and IgM 260 mg/dL (normal 70 to 180 mg/dL). Laboratory studies revealed a normal hemoglobin level, liver enzyme values, white blood cell count, and platelet count but an elevated erythrocyte sedimentation rate (ESR) of 65 mm/h. At this time, T-cell studies were not done. She was treated with antirheumatic drugs with some improvement. Since then the patient has occasionally complained of fever, headache, and arthritis. In September 1989, she suddenly presented with a clear butterfly rash on her face, fever, headache, polyarthritis, mild alopecia, and photosensitivity. At this time she had a high anti-nDNA titer at 1:2,560, ANA titer of 1:1,280, and hypocomplementemia (C3,19 mg/dL; C4,6 mg/dL). Her ESR was 110 mm/h, hemoglobin was 9.2 mg/dL, and white blood cell count was 2.5 X log/L with 24% lymphocytes, 61% segmented neutrophils, 11% band forms, and 4% monocytes. Serum creatinine level, liver enzyme values, and results of urinalysis were normal. There was no serologic evidence of human immunodeficiency virus, hepatitis, Epstein-Barr virus, or cytomegalovirus infection. Liver, spleen, and lymph nodes were not enlarged. There was no evidence of malignancy. She was diagnosed as having SLE as per the American Rheumatism Association’s (ARA) revised criteria [21]. At this time, detailed immunologic studies were performed on blood samples. She was given 80 mgld of prednisolone. Over the next 10 days, her condition deteriorated and she developed nephrotic-range proteinuria followed by thrombocytopenia (platelet count 105 X log/L) and memory disturbances. A computed tomographic brain scan showed multiple infarcts. She died suddenly in the intensive care unit 1 week later from massive brain infarction and septic shock. Patient Control We examined a 29-year-old female Saudi patient with SLE who fulfiied the ARA revised criteria
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[21]. This patient had positive ANA (1:640 titer) and anti-nDNA (1:1,280 titer), decreased complement levels (C3,27 mg/dL; C4,12 mg/dL), and elevated serum IgG, IgA, and IgM levels. The clinical signs included a butterfly rash on the face, mild alopecia, polyarthritis, and thrombocytopenia. T-Cell Studies Peripheral blood mononuclear cells (MNC) were obtained by Histopaque-1077 (Sigma Chemical Co., St. Louis, MO) density gradient centrifugation. B and T cells were separated by the neuraminidasetreated sheep erythrocyte rosetting (E-rosette) technique [22]. CD4+ T cells were purified from Erosetting T cells using OKT8 monoclonal antibody solutions by a panning technique described earlier WI.
Lymphocyte Surface Marker Analysis Surface membrane phenotyping of MNC was determined with the OK and Leu series of monoclonal antibodies (Ortho Diagnostics, Raritan, NJ, and Be&on Dickinson, Mountain View, CA) by the standard indirect immunofluorescence technique [8,22]. The CD nomenclature and specificity of the monoclonal antibodies are shown in Table I. Proliferation and IL-2 Production The proliferative response to anti-CD3 monoclonal antibody (OKT3, 0.2 rg/mL), phytohemagglutinin (PHA, 2 rg/mL), and concanavalin A (Con A, 5 rg/mL) was determined on patient and control T cells as described earlier [7,22]. In brief, 2.5 X lo5 purified E-rosetting T cells were cultured with antiCD3, PHA, or Con A for 72 hours at 37OC in RPMI1640 medium, and incorporation of 3H-thymidine (3H-TdR) into DNA was determined. IL-2 activity defined as U/mL from the supernatants of T cells (2.5 X 105) cultured for 48 hours with mitogens used in the proliferative response was measured in a standard proliferation assay using an IL-2-dependent murine cytotoxic T-lymphocyte cell line as described [22]. B-Cell Growth Factor (BCGF) and B-Cell Differentiation Factor (BCDF) Activity T cells (5 X 105) were incubated in 1 mL of RPMI1640 medium with 1% fetal calf serum, 5 X 10s5 2mercaptoethanol, and 2 rg/mL of PHA. After 72 hours, the supernatants were recovered and were analyzed for BCGF and BCDF activity. BCGF activity from the T-cell supernatants was determined with normal B cells by incorporation of 3H-TdR according to the standard co-stimulator assay [22]. BCDF activity from the T-cell supernatants was
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determined by using Staphylococcus aureus Cowan I (SAC&stimulated normal B cells. B cells were cultured with 0.005% SAC and T-cell supernatanta for 7 days, and were then assayed for IgM and IgG secretion by enzyme-linked immunosorbent assay as shown [22]. Autologous Mixed Lymphocyte Reaction (AMLR) and Allogeneic Mixed Lymphocyte Reaction (MLR) T (92% to 95% E-rosetting and CD2+ cells) and non-T cells (62% to 74% CD24+ B cells, 15% to 18% CDll+ monocytes, and 1% to 4% CD2+ cells) were isolated from MNC by the E-rosetting technique [22]. The non-T cells were pretreated with 50 rg/mL of mitomycin-C (Sigma) for 45 minutes at 37OC. The AMLR and MLR activity of T cells was determined by culturing equal numbers of T cells with mitomycin-C-treated autologous non-T cells (for AMLR) or mitomycin-C-treated allogeneic normal non-T cells (for MLR) for 7 days at 37OC in a proliferation assay [7,22].
RESULTS The surface phenotypes of MNC from two SLE patients, one with deficient (patient) and the other with normal (patient control) TcRc@ and CD3 protein on T cells, were analyzed (Table I). This patient’s T cells lacked expression of both TcRaj3 and CD3 antigens as determined by staining with TcRla/3 (WT31) and OKT3/Leu-4 monoclonal antibodies. This observation of the deficiency of TcRa/3 and CD3 antigen from the patient was confirmed on several occasions at a l-day interval on freshly isolated MNC and E-rosetting purified T cells by using a range of TcR-l@ (WT31), OKT3, and Leu-4 monoclonal antibody concentrations that brightly labeled T cells from the patient control and normal controls. However, both the patient and the patient control had a norhal percentage of the CD2+ and E-resetting+ pan-T cells. They had increased CD4+ and decreased CD8+ T cells. Most patients with SLE we have investigated earlier [7] had an increased percentage of HLA-DR+ (Ia) cells in MNC, which was due to expression of these antigens on T cells. Of interest, HLA-DR+ cells were normal in MNC of our TcR@/CD3-negative patient with SLE, which probably accounts for expression of these antigens on B cells and some monocytes. However, as expected, an increased percentage of HLA-DR+ cells was present in MNC of our patient control. CD4+ CD45R+ (suppressorinducer) cells were decreased with a concomitant increase in CD4+ CDw29+ (helper-inducer) cells in the patient and patient control as compared with normal controls. Further examination of the puri-
TABLE I Phenotypic Characterization of Peripheral Blood Mononuclear Cells % mAbSpecificity Patient Normal Patient Control Control* T-cell receptor(TcR-l&WT31)
0
71
68 + 6
CD1 (OKT6,thymocytes)
2
4
622
CD2 (OKTll,panT)
74
73
7124
CD3cOKT3,panT)
1
70
72 2 4
CD3 (Leu-4,panT)
0
73
73 + 5
CD4 (OKT4, helperT)
61
54
45 + 4
CD8 (OKT8,suppressorT)
12
17
262
CD24(OKB2,Bcells)
11
13
1422
6
5
922
11
43
1223
E-rosettingt (pan-T-cellantigen)
73
72
72 + 4
CD4tCD45Rtt(OKT4,2H4, suppressor-inducer)
16
21
46 k 6
CD4tCDw29tt(OKT4,4B4, helper-inducer)
78
69
41k4
CDll(OKMI,monocytes) HLA-DR (OKlal,
la antigens)
3
4b = moncxlonal antlbody. lormal age-matched controls (n = 4) ? SD. ., neep erytnrocyte rosettmg cells. tmfied CD4t T cells were stained with monoclonal antibodies to CD45R L?H4) and CDw29 84).
fied CD4+ T-cell subsets from the patient with monoclonai antibodies to TcR@ and CD3 protein confirmed negative staining of these subsets for the TcRc@/CD3 protein complex. The function of purified E-rosetting T cells of the patient, patient control, and normal controls in proliferative response and IL-2 secretion is shown in Table II. As shown, the TcR@/CD3-deficient patient’s T cells demonstrated a highly decreased proliferative response to anti-CD3 monoclonal antibody, and a decreased response to PHA and Con A. Similarly, the culture supernatanta of this patient’s T cells produced little IL-2 in response to stimulation with these mitogens. These functional characteristics of T cells from the patient control were higher than in the patient but lower than in normal controls. Upon stimulation with antigens, normal T cells produce various lymphokines such as IL-2, BCGF, and BCDF. The BCGF and BCDF are of crucial importance in humoral immune response and antibody synthesis, which provides a stimulus for differentiation of B cells into antibody-secreting plasma cells. Of interest, although IL-2 activity in the TcRa/3/CD3-deficient patient’s T cells was highly
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TABLE II Functional Characteristicsof T Cells in Proliferative Responseand Interleukin-2 (IL-21 Secretion T Cells
Anti-CD3
PHA
Con A
1.8 + 0.6 6
16.6 + 2.2 21
9.2 k 1.8
38.4 + 4.3 37
55.6 f 4.0 46
32.6 ? 3.1 35
112.4 + 12.3
174.6 ? 17.8
70
93
Patient Proliferation* IL-2 activity+ Patient control Proliferation IL-2 activity Normal control Proliferation IL-2 activity
17
152.6
? 14.5
78
Phfl = pnyronemagglunnln; Mn A = concanavalln A. *Prokferabon (cpm x lo? was determined by culturing T cells with mitogens for 72 hours and incorporatIon of 3H-thymldme, cpm + SD of triplicate cultures after subtracting the background counts. 111-2activity (U/mL) was determined from the supernatants of T cells cultured for 48 hours with mitogens in a proliferationassay usingthe IL-2-dependent cytotoxic T-lymphocyte cell line.
decreased, such cells demonstrated increased BCGF and BCDF activity as compared with that in the patient control’s and normal control’s T cells (Table III). Because of functional and lymphokine abnormalities present in the TcRo$/CD3-deficient patient’s T cells and because of the notion that AMLR and MLR reflect an immunoregulatory mechanism that is important in SLE and other T-cell-mediated diseases, we have also examined the capacity of this patient’s T cells to proliferate in AMLR and MLR. AB shown in Table IV, highly deficient AMLR and MLR activity of the TcR&CD3-deficient patient’s T cells was evident as compared with that in the patient control and normal controls.
COMMENTS We describe a patient with SLE whose peripheral blood T cells expressed CD2+ and E-rosetting panT cells but lacked the normal TcRapKD3 surface protein complex. These TcRa$/CD3 protein complex-negative T cells expressed either the CD4+ or CD8+ phenotype. Expression of both TcRaS and CD3 proteins together on CD4+ or CD8+ subsets is
essential for antigen-specific T-cell function. As a result of deficient cell surface expression of the TcR@3/CD3 complex, T-cell proliferation in response to anti-CD3 monoclonal antibody, PHA, and Con A was impaired, as was IL-2 secretion. Likewise, these T cells showed a highly reduced capacity to proliferate when cultured with either autologous or allogeneic non-T cells in the standard AMLR and MLR assays. However, they were able to secrete elevated levels of two terminal B-cell stimulating factors, BCGF and BCDF. These data suggest that the regulation of IL-2 secretion and BCGF and BCDF secretion by T cells in SLE is independent. On the basis of these findings, we demonstrated that a loss of the TcR@/CD3 protein complex from T cells does not change their ability to influence B-cell function in antibody production, which is a pathologic sequela of T cells in SLE. Investigations of immune function in patients with SLE have revealed IL-2 deficiency and T-cell subset abnormalities [l-8]. In particular, those patients with renal disease and central nervous system involvement have an absolute depression of the CD4+ CD45R+ suppressor-inducer subsets in the circulation [8,23]. The essential findings in both of our SLE patients, the TcRa/.VCD3 protein complex-deficient patient and the patient control, have been depletion of CD4+ CD45R+ cells with an expansion of CD4+ CDw29+ cells. Thus these results parallel those reported earlier by us and others [8,23]. The precise mechanism of the decreased CD4+ CD45R+ cells in SLE is not very clear. However, sera of the patients with a loss of CD4+ CD45R+ cells could have anti-T-cell antibodies directed against this subset, and therefore this may result in their elimination from the blood [4]. Alternatively, the CD4+ CD45R+ cells may represent virgin or naive cells and the CD4+ CDw29+ cells memory [24], and the abnormality observed in SLE may represent a block in differentiation. It is likely that environmental factors such as viruses, ultraviolet light, drugs, or other insults could possibly contribute continuously to these T-cell subset abnor-
TABLE Ill B-Cell Growth Factor (BCGF)and B-Cell Differentiation Factor (BCDF)Activity of T Cells T Cells Stimulated With
Patient BCGF* (cpml
None
1,830
PHA
17,840
BCDF+ Ifi
IN
620 10,200
890 19,500
BCGF (cpm) 1,320 14,290
PatientControl BCDF IgG kM 530 8,760
BCGF (cpm)
NormalControl BCDF M kM
810
1,040
310
15,220
8,300
4,600
iA = phytohemagglutinin. ICGF activity, %thymidine incorporation, meanof triplicate cultures was examinedin a proliferationassay of B cells with T-cell-stimulated culture supernatants Data shown are in cpm. CDFactivity, I& and IgM secretion (ng/mL), was examined by culturing B cells with mitogen-stimulatedT-cell supernatants.
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T CELLS IN SLE / RAZIUDDIN ET AL
malities in the genetically predisposed host. The pathologic significance of this finding may be related directly to enhanced T-cell helper-inducer function by the CD4+ CDw29+ subsets on B-cell hyperactivity in autoantibody secretion, characteristic of this disease. Therefore, the increased secretion of BCGF and BCDF in the T cells of our TcRa/3/CD3 protein complex-negative patient should account for the increased representations of the CD4+ CDw29+ helper-inducer subsets in T-cell populations. The AMLR represents a proliferative response of T cells to self-Ia antigens and that class II major histocompatibility complex gene products alone on autologous non-T cells are capable of triggering this response [6,25]. Moreover, it would appear that the AMLR and MLR in humans are regulated by the same set of rules that govern antigen-specific T-cell activation. We show that the T cells of our TcRc&CD3 protein complex-negative patient did not proliferate either in AMLR or in MLR assays. However, the TcRc@/CD3-positive SLE patient’s T cells demonstrated lower AMLR and MLR activity than in normal controls, but produced higher activity than the TcRaglCD3negative cells. Thus, the variations seen between TcRc@/CD3-negative and positive T cells from the SLE patients suggest that these differences in the functional behavior may be due to the expression of TcR@/CD3 complexes. Genetic predisposition is known to play a crucial role in the development of SLE [26,27]; genetic factors may also contribute to some of the specific abnormalities in T cells. Family studies in the patient’s two healthy children (8-year-old girl and 6year-old boy) and a healthy sister (23 years old) demonstrated that these individuals had a normal percentage of TcR@/CDS-positive T cells in blood (data not shown). There is no family history of immunodeficiency or SLE. Therefore, it is most likely that deficiency of the TcRa@/CD3 protein complex on the T cells of this patient is an acquired abnormality that occurred during the development of SLE. Alternatively, the serum of this patient may have anti-TcRaj3 and anti-CD3 antibodies, which might block or modulate expression of the TcRatflICD3 protein complex on T cells. There are four groups of rearranging genes in human T cells that encode the T-cell receptors for antigens (TcRa, /3, y, and 6). T cells recognize antigens in the context of the major histocompatibility complex through CDS-associated receptors: (~0 chain on mature cells and y6 chain on immature cells [13,14]. Several lines of evidence indicate that expression of both TcR@ and CD3 protein complexes is essential for CD4+ helper/inducer and
TABLE IV Autologous Mixed LymphocyteReaction (AMLR) and Allogeneic Mixed LymphocyteReaction (MLR) Activity of T Cells 1 Cells From Subjects
3H-Thymidine AMLR*
Patient Patlent control Normal control
2.3 + 0.8 28.6 e 3.4 46.5 2 5.2
Incorporation
kpm x 103) MLR+ 1.7 2 0.5 23.2 2 2.8 39.5 2 3.7
I
*AMLR acbvlty was examined by culturing T cells with autologous mitomycln C-treated non-T cells. ‘MLR activity was examined by culturingT cells with mitomycin C-treated allogenelcnormal non-T cells. tData shown are mean r SD of triplicate cultures after subtracting the background counts (T cells cultured alone in medium).
CD8+ suppressor/cytotoxic T-cell function in an immune response [ll-141. Earlier, TcRc@/CD3 protein complex deficiency on T cells was reported from T-lineage lymphomas [16] and in a patient with severe combined immunodeficiency [17]. However, the discordant expression of TcRa@ and CD3 (TcRafl+/CD3-, TcRc+/CD3+) on T cells is a common finding in patients with juvenile rheumatoid arthritis [12], leprosy [ 151, non-Hodgkin’s lymphomas [ 161, Hodgkin’s disease [ 191, and ataxia telangiectasia [20], and has been reported in a patient with combined immunodeficiency with features of graft-versus-host disease [18]. In juvenile rheumatoid arthritis, TcRo$/CD3+ cells represent a notable fraction of spontaneously activated T cells at the site of the lesion in joints, which has led to the speculation that they are involved in the pathogenetic mechanisms occurring in this disease [12]. In conclusion, we have described a new population of human T cells, all with the TcR&CD3phenotype in the circulation of a patient with SLE. On the basis of the monoclonal antibody reactivity of MNC, these TcRo$VCD3- T cells may consist of two distinct subsets, a major population with the TcRaB-, CD3-, CD4+ phenotype and a minor population with the TcRa/3-, CD3-, CD8+ phenotype. The CD4+ populations may further be subdivided into TcR&-, CD3-, CD4+ CD45R+ and TcRafl-, CD3-, CD4+ CDw29+ subsets. Because of our patient’s death, we have not performed studies on T-cell receptor gene rearrangements, and have not determined whether these T cells express an alternative TcR$ chain instead of TcRc@. However, it is unlikely that this TcRaB/CD3T-cell population may express the TcRys chain. The existing evidence that TcRy&positive T cells express neither CD4 nor CD8 antigens lends support to this interpretation. On the basis of functional characteristics observed in proliferative response, AMLR and MLR assays, and lymphokine secretion, it was demonstrated that some usual functions of T cells
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in disease states are severely modified, whereas others are not. We believe that this may be due to the deficiency of TcRcq3 and CD3 on the surface of such cells. Whether such TcR&CD3protein complex-deficient T cells can be identified from immunodeficient cases and additional patients with SLE and other human autoimmune diseases in the future is awaited. In this regard, it will be of interest to monitor changes in the phenotype and pattern of response during the course of the disease. As more is learned about immunoregulatory circuits affecting T-cell receptor gene products in humans, the processes regulating these events in Tcell differentiation pathways may be better understood.
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10. Lanier LL, Federspiel NA, Ruitenberg JJ. The T cell antigen receptor complex expressed on normal peripheral blood CD4-, CD&T lymphocytes: a CD3 associated disulfide linked y chain heterodimer. J Exp Med 1987; 165: 1075-94. 11. Allison JP, Lanier LL. The T cell antigen receptor gamma gene: rearrangement and cell lineage. lmmunol Today 1987; 8: 293-313. 12. DeMaria A, Malnati M. Moretta A, et al. CD3+ 4- B- WT31- (T cell receptor y+) cells and other unusual phenotypes are frequently detected among spontaneously interleukin-2 responsive T lymphocyte present in the joint fluid in juvenile rheumatoid arthritis: a clonal analysis. Eur J lmmunol 1987; 17: 1815-g. 13. Allison JP. Lanier LL. Structure, function and serology of the T cell antigen receptor. Annu Rev lmmunol 1987; 5: 503-40. 14. Strominger JL. Developmental biology of T cell receptors. Science 198% 244: 943-50. 15. Modlin RL. Brenner MB, Krangel MS, Duby AD, Bloom BR. T cell receptors of human suppressor cells. Nature 1987; 329: 541-4. 16. Picker LJ, Brenner MB, Weiss LM, Smith SD, Warnke RA. Discordant expression of CD3 and T cell receptor beta chain antigens in T-lineage lymphomas. Am J Pathol 1987; 129: 434-40. 17. Alarcon B. Requeiro JR, Arnaiz-villena A, Terhorst C. Familial defect in the T cell receptor-CD3 complex. N Engl J Med 1988; 319: 1203-B. 16. Wirt DP. Brooks ED, Vaidys S. et a/. Novel T lymphocyte population in combined immunodeficiency with features of graft-versus host disease. N Engl J Med 1989; 321: 370-4. 19. Dallenbach FE, Stein H. Expression of T cell receptor j3 chain in Reed Sternberg cells. Lancet 1989; 2: 828-30. 20. Carboneri M, Cherchi M, Panganelli P, et al. Relative increase of T cells expressing the gamma/delta rather than the alpha/beta receptors in ataxia telangiectasia. N Engl J Med 1990; 322: 73-6. 21. Tan EM, Cohen AS, Fries JF, eta/. The 1982 revised criteria for the classification of systemic lupus erythematosus. Arthritis Rheum 1982; 25: 1271-7. 22. Raziuddin S, Teklu B. Severe T lymphocyte immunodeficiency associated with hypogammaglobulinemia: defective lymphokine secretion but enhanced autologous mixed lymphocyte reaction. J Clin lmmunol 1989; 9: 448-53. 23. Morimoto C. Steinberg AD, Letvin NL. et a/. A defect of immunoregulatory T cell subsets in systemic lupus erythematosus patients demonstrated with anti2H4 antibody. J Clin Invest 1987; 79: 762-B. 24.Sanders ME, Makgoba MW, Shaw S. Human naive and memory T cells: reinterpretation of helper-inducer and suppressor-inducer subsets. lmmunol Today 1988; 9: 195-9. 25. Weksler ME, Moody CE Jr, Kozak RW. The autologous mixed lymphocyte reaction. Adv lmmunol 1981; 31: 271-312. 26. Block SR. Lokshin MD, Winfield JB. eta/. Immunologic observations on 9 sets of twins either concordant or discordant for systemic lupus erythematosus. Arthritis Rheum 1976; 19: 545-54. 27. Sakane T. Murakawa Y, Suzuki N, et al. Familial occurrence of impaired interleukin-2 activity and increased peripheral blood B cells actively secreting immunoglobulins in systemic lupus erythematosus. Am J Med 1989; 86: 385-90.
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