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347 Regulation of human vascular endothelial cell interleukin-1 production
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ANTIGEN-SPECIFIC SUPPRESSORCELLS IN PATIENTS RECEIVING IMMUNOTHERAPY.Hiroshi Nagaya, M.D., Long Beach, California Previously, we reported that in patients with rye grass hay fever, immunotherapy induces a subpopulation of peripheral blood T cells which are adherent to rye grass antigen(RGA)-coated plates and are less responsive than RGA non-adherent cells when stimulated by RGA. In order to define this subpopulation, peripheral blood T cells were separated by sheep red cell rosetting and fractionated into RGA adherent and non-adherent cells by incubating in RGA-coated plates. The RGA-stimulated proliferative response(S1) of RGA non-adherent cells was reduced from 2.31+0.37 to 1.13+0.25(p
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348 TETANUSTOXOID INDUCES POLYCLONALB CELL ACTIVATION AND RHEUMATOIDFACTORPRODUCTION. L. Tar, M.D. and A.I. Levinson, M.D., Philadelphia, PA Autoantibodies are produced when self-reactive clones are stimulated by well defined polyclonal B cell activators (PBA). Conventional antigens like tetanus toxoid (TT) may elicit polyclonal B cell activation but it is not known if autoantibody production accompanies this response. We analyzed quantitative and qualitative features of the polyclonal immunoglobulin response of peripheral blood mononuclear cells (PBM) stimulated immuin vitro with TT. PBM from 15 previously nized healthy donors were cultured for 9 days with TT (4 ug/ml) or pokeweed mitogen (PWM), a standard PBA. Supernatants were harvested for determination of total IgG, IgM, and IgM rheumatoid factor (IgMRF) by ELISA. TT induced polyclonal IgG and IgM responses in cultures from 7/15 and 6/15 subjects, respectively, whereas PWM induced such responses in 15/15 subjects. TTinduced IgG and IgM responses were lower than those stimulated by PWM. No correlation was observed between 1) TT and PWM-induced IgG responses (r=0.46) and 2) TT and PWM-induced IgM responses (r=0.28), suggesting different mechanOf note, these polyclonal isms of activation. responses were'accompanied by in vitro IgMRF production k2.5 ug/ml) in cultures of 3/7 TT reTT-induced sponders and 5115 PWMresponders. production of polyclonal IgG, IgM and IgMRF was not clearly related temporally to booster immuniThus, stimulation with a conventional zation. antigen, 'IT, may not only induce polyclonal B cell activation but also cause production of autoantibody.
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REGULATION OF HUMAN VASCULAR .--ENDOTHELIAL CELI __.-.-.------5 __INTERLEUKIN-1 PRODUCTION.---- E. Epstr-i~QI&!~~ILii_,~~ Windt,MD,L.J. Rosenwasser,MD,, Boston, MA. --We have previously demonstrated t!~e ahi.! icy of human vascular endothelial cells(H'i'?C)to xt as accessory cells for T cell activat'i~?n. I!"Ki: Were harvested from umbilica:l cords by ;,,)I iagcnase digestion and then grown in culture bit:i endothelial cell growth factor. HVEC were n.:c.J as accessory cells in an adherent cell depIilt<,d mi!ogen induced human T cell proliferation ,xsjay. 7. cell proliferation was markedly decrrasc.4 ~.,ith depletion of adherent cells and was recxxstituted with either monocytes or HVEC as sccesscq cells.HVEC conditioned media(HVECS)was ~1.~0 c;l:,ai-.(e cl< restoring adherent cell depleted 1 cc:1 responses to mitogens, suggesting the possib-iii::; of an Tnterleukin-l(IL-l)like activity.Ilsili.; the elnhancement of lectin induced murine thymocyte or cloned antigen specific T cell proliferation as an assay for IL-1 activity,we were able to demonstrate similar activity in cultured HVEC siqfrnatants. The specificity of this activity l~:ns confirmed with a specific rabbit antiserum tc' human IL-l. Chromatographic analysis of concenrcated HVECS revealed a peak of activity in the 1.0-15 kd fraction,characteristic of typical TL-1 .Wr, have found that constitutive HVEC IL-1 p!.oduction can be augmented by endotoxin stimulaiiiin and under some circumstances by sera from patients with active vasculitis. We are pr4esent.l.y evaluating various agents that may decrease HV!?C 11,-l production. Thus HVEC are capable of providing IL-1 activity for T cell activation and 'ire capable of modulating this activity in rcsp
A LONGITUDINAL IMMUNOLOGICALEVALUATION OF HEMOPHILIAC PATIENTS RECEIVING FACTORVIII CONCENTRATE.R.D. d&hero, M.D., C.B. Deul, M.D., Ph.D., W.A. Andes, M.D., B.E. Eozelka, Ph.D. New Orleans, Louisiana We uerformed a orosoective clinical and immunblogical evaiuation of 30 hemophiliac patients over an average time of one year. No patient developed life threatening opportunistic infection or malignancy; however, the immunologic abnormalities and lymphadenopathy initially present in 9 patients (lymphadenopathy In addition 5 patients, group) persisted. representing 24% of the initial group without lymphadenopathy, developed generalized lymphadenopathy (converter group). One episode of idiopathic thrombocytopenia and one episode of staphlococcal sepsis occurred in this "converter" group. Sixteen patients remained At the time of the follow-up asymptomatic. initial T-lymphocyte subpopulation evaluation, abnormalities persisted, but were not different Natural killer among the three patient groups. cell function alone or in the presence of biologic response modifiers was not different The among hemophiliac and control groups. converter group had significantly better lymphocyte mitogenic function prior to developing lymphadenopathy than the other two hemophiliac groups. Abnormal mitogenle responses present in all patients were explained in part by high levels of spontaneous suppressor cell activity in mononuclear cell preparations. Lymphocyte mitogenic responses of all three hemophiliac groups significantly deteriorated over the course of the study.
346
ANTIGEN-SPECIFIC SUPPRESSORCELLS IN PATIENTS RECEIVING IMMUNOTHERAPY.Hiroshi Nagaya, M.D., Long Beach, California Previously, we reported that in patients with rye grass hay fever, immunotherapy induces a subpopulation of peripheral blood T cells which are adherent to rye grass antigen(RGA)-coated plates and are less responsive than RGA non-adherent cells when stimulated by RGA. In order to define this subpopulation, peripheral blood T cells were separated by sheep red cell rosetting and fractionated into RGA adherent and non-adherent cells by incubating in RGA-coated plates. The RGA-stimulated proliferative response(S1) of RGA non-adherent cells was reduced from 2.31+0.37 to 1.13+0.25(p
347
348 TETANUSTOXOID INDUCES POLYCLONALB CELL ACTIVATION AND RHEUMATOIDFACTORPRODUCTION. L. Tar, M.D. and A.I. Levinson, M.D., Philadelphia, PA Autoantibodies are produced when self-reactive clones are stimulated by well defined polyclonal B cell activators (PBA). Conventional antigens like tetanus toxoid (TT) may elicit polyclonal B cell activation but it is not known if autoantibody production accompanies this response. We analyzed quantitative and qualitative features of the polyclonal immunoglobulin response of peripheral blood mononuclear cells (PBM) stimulated immuin vitro with TT. PBM from 15 previously nized healthy donors were cultured for 9 days with TT (4 ug/ml) or pokeweed mitogen (PWM), a standard PBA. Supernatants were harvested for determination of total IgG, IgM, and IgM rheumatoid factor (IgMRF) by ELISA. TT induced polyclonal IgG and IgM responses in cultures from 7/15 and 6/15 subjects, respectively, whereas PWM induced such responses in 15/15 subjects. TTinduced IgG and IgM responses were lower than those stimulated by PWM. No correlation was observed between 1) TT and PWM-induced IgG responses (r=0.46) and 2) TT and PWM-induced IgM responses (r=0.28), suggesting different mechanOf note, these polyclonal isms of activation. responses were'accompanied by in vitro IgMRF production k2.5 ug/ml) in cultures of 3/7 TT reTT-induced sponders and 5115 PWMresponders. production of polyclonal IgG, IgM and IgMRF was not clearly related temporally to booster immuniThus, stimulation with a conventional zation. antigen, 'IT, may not only induce polyclonal B cell activation but also cause production of autoantibody.
191
REGULATION OF HUMAN VASCULAR .--ENDOTHELIAL CELI __.-.-.------5 __INTERLEUKIN-1 PRODUCTION.---- E. Epstr-i~QI&!~~ILii_,~~ Windt,MD,L.J. Rosenwasser,MD,, Boston, MA. --We have previously demonstrated t!~e ahi.! icy of human vascular endothelial cells(H'i'?C)to xt as accessory cells for T cell activat'i~?n. I!"Ki: Were harvested from umbilica:l cords by ;,,)I iagcnase digestion and then grown in culture bit:i endothelial cell growth factor. HVEC were n.:c.J as accessory cells in an adherent cell depIilt<,d mi!ogen induced human T cell proliferation ,xsjay. 7. cell proliferation was markedly decrrasc.4 ~.,ith depletion of adherent cells and was recxxstituted with either monocytes or HVEC as sccesscq cells.HVEC conditioned media(HVECS)was ~1.~0 c;l:,ai-.(e cl< restoring adherent cell depleted 1 cc:1 responses to mitogens, suggesting the possib-iii::; of an Tnterleukin-l(IL-l)like activity.Ilsili.; the elnhancement of lectin induced murine thymocyte or cloned antigen specific T cell proliferation as an assay for IL-1 activity,we were able to demonstrate similar activity in cultured HVEC siqfrnatants. The specificity of this activity l~:ns confirmed with a specific rabbit antiserum tc' human IL-l. Chromatographic analysis of concenrcated HVECS revealed a peak of activity in the 1.0-15 kd fraction,characteristic of typical TL-1 .Wr, have found that constitutive HVEC IL-1 p!.oduction can be augmented by endotoxin stimulaiiiin and under some circumstances by sera from patients with active vasculitis. We are pr4esent.l.y evaluating various agents that may decrease HV!?C 11,-l production. Thus HVEC are capable of providing IL-1 activity for T cell activation and 'ire capable of modulating this activity in rcsp
A LONGITUDINAL IMMUNOLOGICALEVALUATION OF HEMOPHILIAC PATIENTS RECEIVING FACTORVIII CONCENTRATE.R.D. d&hero, M.D., C.B. Deul, M.D., Ph.D., W.A. Andes, M.D., B.E. Eozelka, Ph.D. New Orleans, Louisiana We uerformed a orosoective clinical and immunblogical evaiuation of 30 hemophiliac patients over an average time of one year. No patient developed life threatening opportunistic infection or malignancy; however, the immunologic abnormalities and lymphadenopathy initially present in 9 patients (lymphadenopathy In addition 5 patients, group) persisted. representing 24% of the initial group without lymphadenopathy, developed generalized lymphadenopathy (converter group). One episode of idiopathic thrombocytopenia and one episode of staphlococcal sepsis occurred in this "converter" group. Sixteen patients remained At the time of the follow-up asymptomatic. initial T-lymphocyte subpopulation evaluation, abnormalities persisted, but were not different Natural killer among the three patient groups. cell function alone or in the presence of biologic response modifiers was not different The among hemophiliac and control groups. converter group had significantly better lymphocyte mitogenic function prior to developing lymphadenopathy than the other two hemophiliac groups. Abnormal mitogenle responses present in all patients were explained in part by high levels of spontaneous suppressor cell activity in mononuclear cell preparations. Lymphocyte mitogenic responses of all three hemophiliac groups significantly deteriorated over the course of the study.