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344 Presence of auto-anti-idiotypic antibodies in house dust mite (HDM) allergic patients
341
PATIENTS
WITH
SELECTIVE
IgA
DEFICIENCY/AT
HAVEANTI HTLV l-MA ANTIBODIES.R.H. Tomall,
343
Bose,Ph.D., D. Marsh, Ph.D., A.H. Sehon, Ph.D., M.D., Ph.D., D.Sc. and G. Delespesse, Winnipeg, Manitoba, Canada, and Baltimore, Md. Auto-ant i-idiotypic antibodies were screened in the sera of 14 individuals who received The sera immunotherapy with Rye grass extract. from serial bleeding obtained over a one year period were tested for (a) Rye I specific IgG and (b) IgG anti-idiotypic and IgE antibodies, antibody specific to IgG F(ab’)* anti-Rye I. IgG F(ab’) anti-Rye I antibody was prepared from the plzsma of one desensitized individual. The sera to be tested for auto-anti-idiotypic antibody were absorbed on Rye I Sepharose-4B and normal human Ig Sepharose-4B in order to any Rye I specific idiotype and remove anti-F(ab’ ) The specificity of activity. the assay if demonstrated by the ability of
D.L. Nelson, P.A. John, and B. Poiesz, Syracuse, New York and Bethesda, Maryland Previouslv we demonstrated that patients and AIDS had increased antibody with pre-AIDS” titers to HTLV l-Membrane antigens (MA). We were concerned that this response might represent another secondary infection. Therefore, we surveyed a group of 60 subjects with primary immunodeficiencies for anti HTLV l-MA. Sera from subjects where obtained from either the National Cancer Institute Metabolism Samples Branch or the Upstate Medical Center. were coded and run together with samples from patients with AIDS as well as controls. Antibody to HTLV l-MA was determined by immunofluorescence cell line (HUT 102 using an HTLV 1 infected 82) as target and analyzed on a flow cytometer. Mean peak fluorescence was measured for each dilution and compared to mean peak fluorescence of negative controls run the same day. A difference of three or more channels was considered positive for that dilution. Seven of 13 (54%) selective IgA deficient subjects and 7 of 18 (39%) ataxia telangiectasia (AT) patients had titers of 61:40. with other primary Only 3 of 29 (10%) patients Previously immunodef iciencies had such levels. we demonstrated that 78 of 108 (72%) AIDS and pre AIDS patients compared to 26 of 339 We believe (8%) controls had titers of 21:40. that these results suggest that HTLV infects a significant proportion of .patients who are deficient in IgA and may contribute to some component of their illness.
ANTI-TETANUS TOXOID (TT) ANTIBODY m A~To-L~NTIIDIOTYPE (Id) ANTIBODY RESPONSES OF YOUNG AND OLD HUMAN SUBJECTS FOLLOWING BOOSTING WITH TETANUS TOXOID. E. Arreaza, M.D., J. Gibbons, Ph.D. ~ M. Weksler, M.D., G. Siskind, M.D. New York, NY employing animal models, In previous studies, we have found that aging is associated with increased production of auto-anti-Id antibodies following primary immunization with haptenIn the present studies,we have conjugates. examined the responses of 3 elderly (>65) and 3 younger (<33) subjects to boosting with TT. AntiTT antibody was assayed by enzyme-linked immunosorbent assay (ELISA) using enzyme-labeled goat Auto-anti-Id antibody was anti-human IgG. assayed by ELISA using the following sequence of additions to polyvinyl microtitre wells: F(ab’)2 fragments of purified human anti-TT; BSA; diluted serum to be assayed; enzyme-labeled goat antihuman Fc; enzyme substrate. Using these assays (1) The the following observations were made. younger subjects produced approximately threefold higher peak titres of IgG anti-TT antibody than the elderly. (2) Peak anti-TT titres occurred at 20 days for the younger and at 30-40 days for the older subjects. (3) There was no difference in the auto-anti-Id responsesofold It is suggested that the and young subjects. failure to observe an increase in auto-anti-Id in the elderly subjects may be because a secondary, rather than a primary response, was studied. (Supported in part by research grant, USPHS, NIH AG 00541; The Lillia Babbitt Hyde Foundation; and The National Cancer Cytology Center).
AUTO-ANTI-IDIOTYPIC ANTIBODYLEVELSIN HAY FEVERPATIENTS TREATEDBY IMMUNOTHERAPY. R.
~P~~i~~‘~;RY~5:_~~~i~~~~‘~2F~~b~~hi~~tI~~~ anti-idiotype; unrelated aff inity2 purifed anti-tetanus IgG) were unable idiotypes (e.g. In all the 14 to inhibit this reaction. individuals studied, auto-anti-idiotype IgG leve 1 changed during desensitization therapy. Complete inverse relationship between idiotype and anti-idiotype levels in 4 individuals suggest that auto-anti-idiotype antibodies may play a role in the modulation of anti-Rye I antibody production.
344
190
PRESENCE OF AUTO-ANTI-IDIOTYPIC AKPIBODIES IN HOUSE DUST MITE (HDM) ALLERGIC PATIENTS. J.-P. Wu, M.D., R. Psuwels, M.D., A.H. Sehon, M.D., Ph.D., and G. Delespesse, Ph.D., D.Sc. Winnipeg, Manitoba, Canada and Gent, Belgium. Thirteen sera from patients allergic to HDM were pooled and extensively adsorbed with HDM Sepharose-4B as well as with IgG- and F(ab') 4 Sepharose in order to remove anti-HD antibodies as well as anti-heavy chain and After absorption, anti-light chain activity. the pool was precipitated twice by 50% ammonium sulfate and fractionated by ACA 34 gel Two peaks were collected, peak I filtration. contained mainly IgM and IgA and peak II Peaks I and II were contained mainly IgG. tested for their ability to block the binding of radiolabelled HDM antigen (purified by gel filtration) on IgE and IgG antibodies from 23 patient sera, different Erom those included in Dose-response studies indicated that the pool. but not peak II nor norm~~5;-\~~ peak I, specifically blocked the binding of to both IgE and IgG antibodies for each sera. The inhibition ranged from 10 to 50% for IgE and from 15 to 30% for IgG antibodies Peak I and II did not interfere ant ibodies. with the binding of an unrelated antigen (Rye when tested I ) to its corresponding antibodies in identical conditions as the anti-HDM shown that peak I It was further ant ibodies. did not contain either IgM, nor IgA anti-HDM, It is therefore nor free HDM antigen. concluded that peak I contains auto-aId crossreacting with public Id expressed on IgE and IgG anti-HDM.
PATIENTS
WITH
SELECTIVE
IgA
DEFICIENCY/AT
HAVEANTI HTLV l-MA ANTIBODIES.R.H. Tomall,
343
Bose,Ph.D., D. Marsh, Ph.D., A.H. Sehon, Ph.D., M.D., Ph.D., D.Sc. and G. Delespesse, Winnipeg, Manitoba, Canada, and Baltimore, Md. Auto-ant i-idiotypic antibodies were screened in the sera of 14 individuals who received The sera immunotherapy with Rye grass extract. from serial bleeding obtained over a one year period were tested for (a) Rye I specific IgG and (b) IgG anti-idiotypic and IgE antibodies, antibody specific to IgG F(ab’)* anti-Rye I. IgG F(ab’) anti-Rye I antibody was prepared from the plzsma of one desensitized individual. The sera to be tested for auto-anti-idiotypic antibody were absorbed on Rye I Sepharose-4B and normal human Ig Sepharose-4B in order to any Rye I specific idiotype and remove anti-F(ab’ ) The specificity of activity. the assay if demonstrated by the ability of
D.L. Nelson, P.A. John, and B. Poiesz, Syracuse, New York and Bethesda, Maryland Previouslv we demonstrated that patients and AIDS had increased antibody with pre-AIDS” titers to HTLV l-Membrane antigens (MA). We were concerned that this response might represent another secondary infection. Therefore, we surveyed a group of 60 subjects with primary immunodeficiencies for anti HTLV l-MA. Sera from subjects where obtained from either the National Cancer Institute Metabolism Samples Branch or the Upstate Medical Center. were coded and run together with samples from patients with AIDS as well as controls. Antibody to HTLV l-MA was determined by immunofluorescence cell line (HUT 102 using an HTLV 1 infected 82) as target and analyzed on a flow cytometer. Mean peak fluorescence was measured for each dilution and compared to mean peak fluorescence of negative controls run the same day. A difference of three or more channels was considered positive for that dilution. Seven of 13 (54%) selective IgA deficient subjects and 7 of 18 (39%) ataxia telangiectasia (AT) patients had titers of 61:40. with other primary Only 3 of 29 (10%) patients Previously immunodef iciencies had such levels. we demonstrated that 78 of 108 (72%) AIDS and pre AIDS patients compared to 26 of 339 We believe (8%) controls had titers of 21:40. that these results suggest that HTLV infects a significant proportion of .patients who are deficient in IgA and may contribute to some component of their illness.
ANTI-TETANUS TOXOID (TT) ANTIBODY m A~To-L~NTIIDIOTYPE (Id) ANTIBODY RESPONSES OF YOUNG AND OLD HUMAN SUBJECTS FOLLOWING BOOSTING WITH TETANUS TOXOID. E. Arreaza, M.D., J. Gibbons, Ph.D. ~ M. Weksler, M.D., G. Siskind, M.D. New York, NY employing animal models, In previous studies, we have found that aging is associated with increased production of auto-anti-Id antibodies following primary immunization with haptenIn the present studies,we have conjugates. examined the responses of 3 elderly (>65) and 3 younger (<33) subjects to boosting with TT. AntiTT antibody was assayed by enzyme-linked immunosorbent assay (ELISA) using enzyme-labeled goat Auto-anti-Id antibody was anti-human IgG. assayed by ELISA using the following sequence of additions to polyvinyl microtitre wells: F(ab’)2 fragments of purified human anti-TT; BSA; diluted serum to be assayed; enzyme-labeled goat antihuman Fc; enzyme substrate. Using these assays (1) The the following observations were made. younger subjects produced approximately threefold higher peak titres of IgG anti-TT antibody than the elderly. (2) Peak anti-TT titres occurred at 20 days for the younger and at 30-40 days for the older subjects. (3) There was no difference in the auto-anti-Id responsesofold It is suggested that the and young subjects. failure to observe an increase in auto-anti-Id in the elderly subjects may be because a secondary, rather than a primary response, was studied. (Supported in part by research grant, USPHS, NIH AG 00541; The Lillia Babbitt Hyde Foundation; and The National Cancer Cytology Center).
AUTO-ANTI-IDIOTYPIC ANTIBODYLEVELSIN HAY FEVERPATIENTS TREATEDBY IMMUNOTHERAPY. R.
~P~~i~~‘~;RY~5:_~~~i~~~~‘~2F~~b~~hi~~tI~~~ anti-idiotype; unrelated aff inity2 purifed anti-tetanus IgG) were unable idiotypes (e.g. In all the 14 to inhibit this reaction. individuals studied, auto-anti-idiotype IgG leve 1 changed during desensitization therapy. Complete inverse relationship between idiotype and anti-idiotype levels in 4 individuals suggest that auto-anti-idiotype antibodies may play a role in the modulation of anti-Rye I antibody production.
344
190
PRESENCE OF AUTO-ANTI-IDIOTYPIC AKPIBODIES IN HOUSE DUST MITE (HDM) ALLERGIC PATIENTS. J.-P. Wu, M.D., R. Psuwels, M.D., A.H. Sehon, M.D., Ph.D., and G. Delespesse, Ph.D., D.Sc. Winnipeg, Manitoba, Canada and Gent, Belgium. Thirteen sera from patients allergic to HDM were pooled and extensively adsorbed with HDM Sepharose-4B as well as with IgG- and F(ab') 4 Sepharose in order to remove anti-HD antibodies as well as anti-heavy chain and After absorption, anti-light chain activity. the pool was precipitated twice by 50% ammonium sulfate and fractionated by ACA 34 gel Two peaks were collected, peak I filtration. contained mainly IgM and IgA and peak II Peaks I and II were contained mainly IgG. tested for their ability to block the binding of radiolabelled HDM antigen (purified by gel filtration) on IgE and IgG antibodies from 23 patient sera, different Erom those included in Dose-response studies indicated that the pool. but not peak II nor norm~~5;-\~~ peak I, specifically blocked the binding of to both IgE and IgG antibodies for each sera. The inhibition ranged from 10 to 50% for IgE and from 15 to 30% for IgG antibodies Peak I and II did not interfere ant ibodies. with the binding of an unrelated antigen (Rye when tested I ) to its corresponding antibodies in identical conditions as the anti-HDM shown that peak I It was further ant ibodies. did not contain either IgM, nor IgA anti-HDM, It is therefore nor free HDM antigen. concluded that peak I contains auto-aId crossreacting with public Id expressed on IgE and IgG anti-HDM.