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708 Modulation of cytokine gene expression by neuropeptides and human immuno-deficiency virus (HIV)peptides
705
706
EFFECTS OF INTERFERON-GAtMA (IFN-Y) ON SURFACE ANTIGEN EXPRESSION IN CHILDREN WITH CHRONIC GRANULOEIATO/SDISE;S;i(CGD). HI4 Rosenblatt. MO, W. Smith, P D. N. a kh. H. Dombrowski, DC. Anderson. MD, Houston, Texas. Two-color flow cvtometrv was used to studv lymphocyte surface markers-in 8 CGD patientsparticipating in a multi-center placebo controlled double blind trial of IFN-‘I. IFN treated (I) and placebo treated (P) groups had significantly lower percentages of CD3t and CD4t T-cells and higher percentages of mature (CD19t) and "activated" (Leu8tCD20t) B-cells than normal adult controls (C). The P and I groups did not differ from each other for these markers. Expression of Leu7, CD8, and CD25 were not for the 3 groups. The mean % + 1 S.D. different of lymphocytes staining for each marker was: Marker C I’ I-!!! 61+10** 58+3 CD3t 6727 31+4 44+7 32+ 8 CD4+ 3125 285 4 CD8+ 32+6 21+ 7 24+4 CD19t 9+3 1953 192 7 Leu8tCD20t 9+4 65+5# 83+6 75+10 CD2t CDIlb 15 1 * pc.iOl; fEi+a:l P an: t3ts contt$C) ** ~1.01 P v. C; # pL.01, ## p(.OOI for I vs P The I group compared with the P group had lower expression of the adhesion molecules CDllb and CD2 (p <.OOI and p c.01 respectively). Alterations of CDllb and CD2 expression may be due either to direct effects of IFN-y on peptide synthesis or to lower infection rates in the treated patients. Modulated expression of adhesion molecules on lymphoid and phagocytic cells may explain some benefits of IFN-yin CGD.
707
RENALFAlLUREASANEWAS6CClATlON WITHANDA POSSIBLE CAUSE OF ACQUIRED LOW SERUM IgG. &jg MD. ;md
708
MeSanDiqjoandLosAngelea,Califomla. Oliguric renal failure (RF) does not cause a reduction in mean serum lgG levels. RF k associated with an increased incidence of infectbns and a number of immune system abnormalities. We have discovered that a subgroup of RF patients on dialysis pre-transplant have very bw serum IgG levels. As part of an unrelated study. all IgG levels <300 mg/dl in individuals >3 years old obtained at UCLA from 1984-87 were identified by retrospective survey. IgG levels were from both in and out patient care and would have been obtained durlng the workup of severe infections. 63 individuals were IdenMecl. The medical records were then reviewed and underlying pathologic processes potentially causing the low IgG levels identified. As expected there were individuals (#) with common variable immunodeflclency (CVI) (14), paraproteins (12). leukemias and lymphomas (ll), lupus with RF (3), chemotherapy (2), congestive heart failure (2), phenyltoin use (l), ataxia telangetasia (1). partially treated SCID (1). GI loss (1) and thrombocytopenia (1). The unexpected flndlng was that 22% of patients had no other known palhobgk process except oligurk RF (14) on dialysis before renal transplant. A prospective blinded study of IgG levels in 46 sequential unselected RF patient sera collected for pre-transplant hepatitis screening was then done. It revealed 3 patients or 6% had unexpected serum IgG ~400 mgldl. The mean serum IgG of the group was (MSD) 1461*716 mg/dl, In the normal range. The possibility of occult but coexistent disease and the prospective natural history of bw IgG in this population needs to be determined. A trial of IVlg to reduce mo&klity in this potentially large group of individuals may be warranted.
Y
QJ?X?
,-J&Cl-S. Schoolev. and B.D.
J.I. V&&&L
Uonn, T.J. Boston
FROM HIVCurie1.11
%A
We utilized an anti-CD3:ant.i-i’08:!.:.‘~.::: .:I .I:, anti-CD3:anti-CD4(CD3,4) bispecifir wAb(RSMAB: system to propagate preferentialli ivi:I-0 ‘kc CD4+ and CD8+ T cell subsets, respnr’ ;\.‘(‘I v -!‘*a::! the peripheral blood of HIV- infecrtad ,.ut, jc<.* b CD3,A causes selective cytolysis .,! .!:r :.:>Xt ! cells by bridging the CD8 molecule:; r,! :-argc’ cells to the CD3 complex of CT1.s :.?i I’:uK:‘:::-~~:! activation and proliferation of !?e :-t,sidu.r! CD3+CD4+ T cells. Similarly. CD3, . ,‘,‘LsE’:; ~r’.cct-ive lysis of CD4t T cells and consur~en~ wr-i‘lation of thr residual CD3+CD8+ T rr:‘~: ir; -,I : subjects tested, CD3.8 expanded rile Cwi slibi.i.[ lo-75 folds and enriched the CD4+ a.-.lIs fror~ au initial range of 3.29% to a final !-~nge o: ::i 95%(<5%CDB+). In contrast., CD7 ..‘a t~qxx~led r!... CD8+ subset 8->25 folds and enriched rh, r‘DR. cells to 80.97%(<3%CD4+). These cul illres ve:e assayed for cytolytic activities ignillsc $1 .d~nel of autologous EBV-transformed lymphob:asrs !IML expressed either the HIV envelope:EF1:, TP‘:F,.s~ cranscriptase(RT). or gag protein “Lr, recolllb;nant vaccinia vectors. 2,/2 CD41 culturcts exhibi.rci. MHC-restricted lysis against ENV- IU: nor ag.ainst RT- or gag- expressing targets ‘i.‘4 c!at cul rurcs assayed against the same panel rxhihired MHC r~stricted lysis agains: RT- and gag- b11r nof against ENV- expressing targets iii conclusio:,, BSMAR provided elegant, rapid means [o nbt-ainrd CD4+ and CD8+ T cell subsets from HIV-infrrt,,;l subjects. The enriched populations thou obtained have high anti -viral activities and rlr~, of r,l-.~ar value in the srxdy of viral antigwic;ry
MODULATION OF CYTOKINE GENE EXPRESSION BY NEUROPEPTIDES AND HUMAN IMMUNODEFICIENCY VIRUS (HIV)PEFTIDES. w A.AnnArbor,SA.. We postulate that depression or disruptive life events following HIV infection rwy activate hypothalmos-pituitaryadrenal axis (HPA) resulting in elevated levels of neuropeptides/hormones which alone or in synergy with HIV derived soluble products can e%aarbatt OTcausepmgressive immunodeficiency and encepbalopathy. Nat& k&r cell activity and the production of various immunoregulatory cytokines are known to be affected by HIV infection. Our previous studies have shown that cOmcosteroids and HIV envelope peptides could induce immunosuppression which could be subsequently reversed by interkukin-2 (IL2). The in vitro effects of neuqeptide Y (NPY) and env 647-659, a synthetic peptide derived from the HIV genome, were examined on IL2, IL2 receptor and interferon m gene expression by normal lymphocytes. Peripheral blood mononuclear cells from healthy adult donors were cultured in media alone or with 5 pg of PHA or env 647-659 (10 @ml) or NPY (10-7M) or PHA + 647-659/NPY. RNA was isolated and 5, 2.5 and 1.25 lg of RNA samples were slot blotted and hybridized with Q2-labeled cDNA probes specific for IL2, ILZR and IFN?R In slot blot analysis, NPY and env 647-659 significantiy suppressed PHA induced mRNA specific for 1L2, ILZR and IFNTR. In northern blot analysis, mRNA specific for IL2 (0.91kb) and IFNm (1.75kb) were detected and NPY and env 647-659 significantly inhibited mRNA specific for IL2 and IFNrR. These studies reveal an immunomcduIatory effect of NPY and HIV peptides on IL2, IL2lR and IFFY ene expression and may lead to a be= understanding o B the pathogenic mcchaniJms underlying the immunodetiency and CNS dysfunction associatedwith HIV infection.
706
EFFECTS OF INTERFERON-GAtMA (IFN-Y) ON SURFACE ANTIGEN EXPRESSION IN CHILDREN WITH CHRONIC GRANULOEIATO/SDISE;S;i(CGD). HI4 Rosenblatt. MO, W. Smith, P D. N. a kh. H. Dombrowski, DC. Anderson. MD, Houston, Texas. Two-color flow cvtometrv was used to studv lymphocyte surface markers-in 8 CGD patientsparticipating in a multi-center placebo controlled double blind trial of IFN-‘I. IFN treated (I) and placebo treated (P) groups had significantly lower percentages of CD3t and CD4t T-cells and higher percentages of mature (CD19t) and "activated" (Leu8tCD20t) B-cells than normal adult controls (C). The P and I groups did not differ from each other for these markers. Expression of Leu7, CD8, and CD25 were not for the 3 groups. The mean % + 1 S.D. different of lymphocytes staining for each marker was: Marker C I’ I-!!! 61+10** 58+3 CD3t 6727 31+4 44+7 32+ 8 CD4+ 3125 285 4 CD8+ 32+6 21+ 7 24+4 CD19t 9+3 1953 192 7 Leu8tCD20t 9+4 65+5# 83+6 75+10 CD2t CDIlb 15 1 * pc.iOl; fEi+a:l P an: t3ts contt$C) ** ~1.01 P v. C; # pL.01, ## p(.OOI for I vs P The I group compared with the P group had lower expression of the adhesion molecules CDllb and CD2 (p <.OOI and p c.01 respectively). Alterations of CDllb and CD2 expression may be due either to direct effects of IFN-y on peptide synthesis or to lower infection rates in the treated patients. Modulated expression of adhesion molecules on lymphoid and phagocytic cells may explain some benefits of IFN-yin CGD.
707
RENALFAlLUREASANEWAS6CClATlON WITHANDA POSSIBLE CAUSE OF ACQUIRED LOW SERUM IgG. &jg MD. ;md
708
MeSanDiqjoandLosAngelea,Califomla. Oliguric renal failure (RF) does not cause a reduction in mean serum lgG levels. RF k associated with an increased incidence of infectbns and a number of immune system abnormalities. We have discovered that a subgroup of RF patients on dialysis pre-transplant have very bw serum IgG levels. As part of an unrelated study. all IgG levels <300 mg/dl in individuals >3 years old obtained at UCLA from 1984-87 were identified by retrospective survey. IgG levels were from both in and out patient care and would have been obtained durlng the workup of severe infections. 63 individuals were IdenMecl. The medical records were then reviewed and underlying pathologic processes potentially causing the low IgG levels identified. As expected there were individuals (#) with common variable immunodeflclency (CVI) (14), paraproteins (12). leukemias and lymphomas (ll), lupus with RF (3), chemotherapy (2), congestive heart failure (2), phenyltoin use (l), ataxia telangetasia (1). partially treated SCID (1). GI loss (1) and thrombocytopenia (1). The unexpected flndlng was that 22% of patients had no other known palhobgk process except oligurk RF (14) on dialysis before renal transplant. A prospective blinded study of IgG levels in 46 sequential unselected RF patient sera collected for pre-transplant hepatitis screening was then done. It revealed 3 patients or 6% had unexpected serum IgG ~400 mgldl. The mean serum IgG of the group was (MSD) 1461*716 mg/dl, In the normal range. The possibility of occult but coexistent disease and the prospective natural history of bw IgG in this population needs to be determined. A trial of IVlg to reduce mo&klity in this potentially large group of individuals may be warranted.
Y
QJ?X?
,-J&Cl-S. Schoolev. and B.D.
J.I. V&&&L
Uonn, T.J. Boston
FROM HIVCurie1.11
%A
We utilized an anti-CD3:ant.i-i’08:!.:.‘~.::: .:I .I:, anti-CD3:anti-CD4(CD3,4) bispecifir wAb(RSMAB: system to propagate preferentialli ivi:I-0 ‘kc CD4+ and CD8+ T cell subsets, respnr’ ;\.‘(‘I v -!‘*a::! the peripheral blood of HIV- infecrtad ,.ut, jc<.* b CD3,A causes selective cytolysis .,! .!:r :.:>Xt ! cells by bridging the CD8 molecule:; r,! :-argc’ cells to the CD3 complex of CT1.s :.?i I’:uK:‘:::-~~:! activation and proliferation of !?e :-t,sidu.r! CD3+CD4+ T cells. Similarly. CD3, . ,‘,‘LsE’:; ~r’.cct-ive lysis of CD4t T cells and consur~en~ wr-i‘lation of thr residual CD3+CD8+ T rr:‘~: ir; -,I : subjects tested, CD3.8 expanded rile Cwi slibi.i.[ lo-75 folds and enriched the CD4+ a.-.lIs fror~ au initial range of 3.29% to a final !-~nge o: ::i 95%(<5%CDB+). In contrast., CD7 ..‘a t~qxx~led r!... CD8+ subset 8->25 folds and enriched rh, r‘DR. cells to 80.97%(<3%CD4+). These cul illres ve:e assayed for cytolytic activities ignillsc $1 .d~nel of autologous EBV-transformed lymphob:asrs !IML expressed either the HIV envelope:EF1:, TP‘:F,.s~ cranscriptase(RT). or gag protein “Lr, recolllb;nant vaccinia vectors. 2,/2 CD41 culturcts exhibi.rci. MHC-restricted lysis against ENV- IU: nor ag.ainst RT- or gag- expressing targets ‘i.‘4 c!at cul rurcs assayed against the same panel rxhihired MHC r~stricted lysis agains: RT- and gag- b11r nof against ENV- expressing targets iii conclusio:,, BSMAR provided elegant, rapid means [o nbt-ainrd CD4+ and CD8+ T cell subsets from HIV-infrrt,,;l subjects. The enriched populations thou obtained have high anti -viral activities and rlr~, of r,l-.~ar value in the srxdy of viral antigwic;ry
MODULATION OF CYTOKINE GENE EXPRESSION BY NEUROPEPTIDES AND HUMAN IMMUNODEFICIENCY VIRUS (HIV)PEFTIDES. w A.AnnArbor,SA.. We postulate that depression or disruptive life events following HIV infection rwy activate hypothalmos-pituitaryadrenal axis (HPA) resulting in elevated levels of neuropeptides/hormones which alone or in synergy with HIV derived soluble products can e%aarbatt OTcausepmgressive immunodeficiency and encepbalopathy. Nat& k&r cell activity and the production of various immunoregulatory cytokines are known to be affected by HIV infection. Our previous studies have shown that cOmcosteroids and HIV envelope peptides could induce immunosuppression which could be subsequently reversed by interkukin-2 (IL2). The in vitro effects of neuqeptide Y (NPY) and env 647-659, a synthetic peptide derived from the HIV genome, were examined on IL2, IL2 receptor and interferon m gene expression by normal lymphocytes. Peripheral blood mononuclear cells from healthy adult donors were cultured in media alone or with 5 pg of PHA or env 647-659 (10 @ml) or NPY (10-7M) or PHA + 647-659/NPY. RNA was isolated and 5, 2.5 and 1.25 lg of RNA samples were slot blotted and hybridized with Q2-labeled cDNA probes specific for IL2, ILZR and IFN?R In slot blot analysis, NPY and env 647-659 significantiy suppressed PHA induced mRNA specific for 1L2, ILZR and IFNTR. In northern blot analysis, mRNA specific for IL2 (0.91kb) and IFNm (1.75kb) were detected and NPY and env 647-659 significantly inhibited mRNA specific for IL2 and IFNrR. These studies reveal an immunomcduIatory effect of NPY and HIV peptides on IL2, IL2lR and IFFY ene expression and may lead to a be= understanding o B the pathogenic mcchaniJms underlying the immunodetiency and CNS dysfunction associatedwith HIV infection.