Serological interactions among sera of human renal graft recipients in double diffusion in gel were observed by chance. Antigen was detected in six of 127 recipients and antibody in two of 30 recipients tested. One of the six recipients carrying the antigen had also antibody in one serum sample. On the basis of the pattern of reactions observed, the hypothesis was expressed that the described antigen-antibody system had "pan" rather than "allo" character.
Antibodies raised in rabbits against pig kidney DOPA decarboxylase show immunological cross-reactivity towards extracts from monkey, beef, rat and rabbit kidney. The influence of the immuno-reaction on the enzymatic activity has been investigated.
Modification of graft immunogenicity using graft (GPTX) and donor pretreatment (DPTX) has been pursued in an attempt to modify allograft immunogenicity using various immunosuppressive agents. The murine skin allograft and canine renal allograft models were used to study the efficacy of Cyclosporine (Cy A) as a DPTX and GPTX prior to transplantation. Tail skin allografts from C3HHeN male mice were grafted to Balb/c female mouse recipients. Minimal immunosuppression was given to all skin graft recipients. Skin allograft were either GPTX with Cy A, DPTX with either Cy A, methylprednisolone (MP), or cyclophosphamide (CP), or Cy A GPTX and DPTX with the three drugs alone or in combination. Cy A GPTX alone of skin allografts did not significantly prolong survival. DPTX with Cy A significantly prolonged skin graft survival, however, CP or MP alone did not. The various combinations of MP, Cy A, and CP as DPTX and MP, Cy A, and CP DPTX used together with Cy A GPTX also significantly prolonged murine skin allograft survival. Kidney allografts used unrelated mongrel dogs as donors or recipients. Renal transplant experimental groups were either: Non-pretreated and immediately transplanted, nonpretreated and hypothermically stored (HS) for 24 hours in Collins (C-2) solution, GPTX with 12.5 mg Cy A during 24 hr. HS in C-2, DPTX with Cy A (25 mg/Kg), or Cy A DPTX (15 mg/kg) and GPTX during 24 hrs. HS in C-2. Cy A GPTX during HS was sometimes effective in prolonging kidney allograft survival greater than 30 days using only minimal immunosuppression with azathioprine. Cy A DPTX prolonged survival somewhat, but not significantly. Improved results were seen, however, when Cy A DPTX was used together with Cy A graft pretreatment. These results indicate the potential for the successful use of Cy A as a donor and/or graft pretreatment, however, further studies will be necessary to optimize the use of Cy A in these modalities.
New microspheres having functional aldehyde groups have been prepared by radiation polymerization of acrolein solution containing hydroxyethyl methacrylate and glutalardehyde. The size distribution in the microspheres was narrow and average particle diameter was 1 - 2 micron. The binding ability of the microspheres to antigen increased by increasing the concentration of glutalardehyde. The preparation procedure of the microspheres is simple. The microspheres can be used for immunoresearch.
Functional differences between adherent macrophages and macrophages in suspension have been repeatedly reported. During the course of developing serum-free phagocytic assays to evaluate Fc gamma receptor (Fc gamma R)-mediated phagocytosis by rat alveolar macrophages (AM) we found Fc gamma R-mediated phagocytosis by Rat AM in suspension was markedly greater than when these cells were bound to plastic substrate in protein-free medium. This dissimilarity in performance was related, in part, to a plastic substrate-associated loss in Fc gamma R-mediated phagocytosis in that this cell function was preserved when monolayers of AM were formed in culture medium containing a low concentration of bovine serum albumin (BSA). The results of particle binding studies suggest the diminished phagocytic activities of AM in monolayers formed in the absence of BSA was due to a loss in the functional expression of Fc gamma R.
Guinea pig heart was perfused in vitro by xenogeneic sera. Sera from both Forssman-positive and Forssman-negative animals exerted strong cardiotoxicity. The toxic effect of rabbit sera could be very significantly decreased by absorption or neutralization of the naturally occurring Forssman antibodies, pointing to the significant role played by these antibodies in the “rejection” of a Forssman-positive organ.
Infectious mononucleosis sera gave positive results in enzymoimmunoassay with glutaraldehyde-treated human erythrocytes. This unexpected reaction appeared to be caused by the interaction of Paul-Bunnell (P-B) antibodies with a partial P-B antigen that apparently appears on human red blood cells in a hidden form and becomes exposed by the treatment with glutaraldehyde.
Lymphoma cells (Eb and ESb, the metastasizing variant) were injected via a mesenteric vein. 85% to 95% of these cells were trapped in the liver. Most of the Eb cells (81%) were destroyed within 18 hours, ESb cells prove to be more resistant: only 28% of these cells were destroyed within this time. 51-Chromium release assay (4 hours) revealed a similar sensitivity of these two tumor cell strains.
VOl.1: Membranes, Detergents and Receptor Solvbilization vol.2: Receptor Purification Procedures Vol.3: Molecular and Chemical Characteristics of Membrane Receptors Alan R. Liss, New York, 1984, hardbound; Vol.1: 238 pages, $46.00; Vol.2: 306 pages, $52.00; Vol.3: 184 pages, $34.00
Aids ib developing into one o t the most instructive imunological diseases since the discovery of monoclonal gammopathies. AIDS studies contiriue to reveal new aspects about the cellularimuuue system and other research triggered by AIDS has led to more than a doubling of the known varieties of veneral diseabe, if that still is the appropriate term. that it is hard t o keep abrecst of the literature, and even books such as tkis one become obsolete the moment they appear i × i print. Nevertheless, as an overview of the field in all its ramifications, as off early 1983, this work is a useful and important docunent. be the ultinate culprit, but in the meantime this collectlon already comprises all the likely candidates, as well as all the auxiliary and secondary pathogens that characterize and/or accompany the syndrome. However, the field moves so fast We'll find out liiter which virus finall) will prove to
A one step solid phase radioimmunoassay is used as a simple and reproducible method of detection and quantitation of IgG produced by human PBL after stimulation with PWM. Modifications of culture conditions are necessary to make culture supernatants suitable for this assay. Pulsing with PWM must be performed in serum-supplemented culture medium for 4-5 days. After thorough washing, medium is then replaced with serum-free medium. Under these conditions, synthesis and secretion of IgG continues for at least 9 days. The amount of IgG produced by 10(6) normal adult PBL as detected in this system is 0.77 +/- 0.47 micrograms. No close correlation between cell proliferation and IgG synthesis was observed.
We have observed that CT6 cell line, a murine interleukin 2 (IL 2)-dependent T cell line, was highly responsive to phorbol myristate acetate (PMA) and to a lesser extent but significantly responsive to lipopolysaccharide (LPS) in the short-term proliferation assay. In contrast, two other murine IL 2-dependent cell lines, CTLL-2 and NK clone 7, were totally unresponsive to these stimulants. Even when CT6 was recloned by a limiting dilution, no unresponsive clone to PMA was obtained, whereas several clones unresponsive to LPS were obtained. PMA, unlike to IL 2, could not support a long-term culture of CT6. There are no differences between CT6 and CTLL-2 except that the former possessed asialo GM1 while the latter lacked it. Though it is unknown whether this difference is involved in the mechanism of the response of CT6 to PMA or LPS, these data give caution to us against the use of CT6 for the determination of IL 2 activity contaminated with PMA and LPS.
The development of a quantitative spreading assay of macrophage activation is described. The assay involved incubation of macrophages on glass coverslips for 1 hour and assessment of cell size using a microscope attached to a microcomputer-assisted digitising system which allowed the diameter of 200 cells to be assessed within 10 minutes. Mouse peritoneal macrophages were used in the development of the assay. Internal consistency of the assay was shown by minimal inter-observer, intra-observer and inter-animal variation. Validation of the assay as a measure of macrophage activation was confirmed by the use of in vivo and in vitro activating agents. Once validated the assay was used to detect activation in alveolar macrophages from rats exposed to airborne asbestos. The macrophage spreading assay described here is quick, reliable, consistent and easy to perform and has a potentially wide application in studies of macrophage function and dysfunction.
Circulating immune complexes were detected by the immunoelectrophoretic method in 18 of 29 (62 per cent) of patients with systemic scleroderma. The presence of immune complexes did not correlate with that of antinuclear antibodies to dsDNA, DNP, RNP, and Sm. The mean levels of immunoglobulins G, A, and M as well as of C3 were significantly higher in patients with systemic scleroderma than in blood donors.
Cerebrospinal fluid (CSF) lymphocytes of 6 multiple sclerosis (MS) patients were cultured with tetanus toxoid (TT) and irradiated autologous antigen presenting cells (APC) followed by propagation of the responding T-cells in interleukin-2 containing medium. TT-reactive cell lines were recovered from 4 of the 6 CSF samples, even though the patients had not been TT booster immunized in recent years. These findings suggest an active circulation of antigen reactive lymphocytes from the systemic immune compartment(s) into the CSF even without recent activation by booster immunization. Since immune reactions to TT are very unlikely to be pathogenic in MS, these findings also indicate that presence of CSF lymphocytes reactive to a particular antigen does not necessarily imply a causal role.
Rabbits immunized with purified Sm and RNP small nuclear ribonucleoproteins (snRNPs) produced precipitating and hemagglutinating antibodies against these antigens. These antibodies had immunological specificity identical to the naturally occurring SLE anti-Sm/RNP antibodies as demonstrated by immunoprecipitation results and studies involving the characterization of immunoaffinity purified antigens isolated from the rabbit immune and SLE anti-Sm/RNP IgG affinity columns.
A micromethod is described which allows subpopulation classification of proliferating (3H thymidine incorporating) cells, previously stimulated in microcultures. The technique is based on transferring 1,000 to 5,000 cells from microcultures to poly-L-lysine coated multispot slides. The cells are then stained for surface markers using the immuno-peroxidase method, combined with subsequent autoradiography.
The oxidative response of murine spleen cells to secondary exposure to antigen was determined by luminol (5-amino-2,3-dihydro-1,4-pthalazinedione) amplified chemiluminescence, CL. BALB/cj and CBA/J mice were immunized with saline or an antigen solution of saline, luminol, and bovine serum albumin. Spleen cells were obtained from mice two and four days after immunization, and the CL response to in vitro antigenic exposure was measured for 35 minutes. At two days post-immunization, there was no difference in the CL of control and antigen-primed cells. By day four, the antigen-primed CL response differed significantly in both magnitude and time course from the primary antigen-stimulated response of the controls. This early development of differential CL response to antigenic challenge suggests a role for oxidative metabolic activity in the expression of the anamnestic immune response.
Rabbit antisera to bovine nerve preparations were used to study the tissue distribution in the ox of P2, an antigen specific for the peripheral nervous system. Double diffusion gel precipitation tests were able to demonstrate P2 in spinal nerves, trigeminal nerve, spinal cord, medulla oblongata, and pons, but not in higher centers of the CNS, optic nerve, or non-neural tissues. A highly sensitive inhibition of enzyme immunoassay was developed to detect and quantitate low levels of P2. By this assay, P2 was found to be most concentrated in peripheral nerves, with decreasing amounts found in the spinal cord, medulla oblongata, pons, cerebellum, and cerebral peduncle. No P2 was found in the thalamus, cerebrum, or optic nerve; however, low levels of P2 were detected in the adrenal medulla, a non-neural tissue composed largely of cells derived from the embryonic neural crest region.
