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Biomedical subjects

C R Parish

Publications and source records attributed to C R Parish.

At least 127 records · Page 7Linked to original sources
PubMed1981

A sensitive and quantitative fluorescence assay for cell surface antigens.

A procedure is described for the assay of cell surface antigens based on quantitative fluorometry. Fluorescent immunospheres are coupled with sheep anti-mouse immunoglobulins or Protein A and used to detect specific antibody bound to target cells. The fluorescent sphere assay described here offers 16--128-fold greater sensitivity than complement mediated lysis or Protein A radioimmune assays and comparable sensitivity to rosetting assays. In addition, the assay is simple to perform, uses commercially available reagents and is completely objective in that a common laboratory fluorometer is used to obtain fluorescence measurements.

Animals↗
PubMed1981

Reactivity of xenogeneic antihuman Ia antisera with the I-C subregion of the murine major histocompatibility complex. I. Genetic studies.

Several rabbit xenoantisera raised against human serum Ia antigens have been shown to detect murine lymphocyte antigens. The murine system detected was shown to be polymorphic and expressed on B cells and not T cells. Serological analysis of recombinant H-2 strains demonstrated that two of the antisera recognized products of the I-Cd subregion, and one antiserum recognized products of the I-Cb subregion, thus clearly establishing the existence of the I-C subregion in these two haplotypes. However, as the I-Cd subregion is classically described by the Ia.6 specificity present on T cells and xenoantisera detect a product on B cells, the indication is that the I-C region, like I-A and I-J, complex and contains genes coding for specificities on T cells or B cells.

Absorption↗
PubMed1981

Anti-self receptors. III. Lack of allelic exclusion and thymic epithelium dependence of H-2L-region-restricted receptors on lymphocytes.

Murine thymocytes and peripheral lymphocytes bind autologous erythrocytes via H-2L-region-restricted receptors. After inhibiting autorosetting with different erythrocyte sonicates the specificity of these anti-self receptors was examined in F1 hybrid and chimaeric mice. Most F1 lymphocytes simultaneously expressed receptors against both parental haplotypes. Furthermore, analysis of lymphocytes from allogeneic and semi-allogeneic chimaeras clearly demonstrated that radioresistant elements in the recipient thymus did not modify the haplotype specificity of the receptors on donor-derived lymphocytes.

Alleles↗
PubMed1981

Serum Ia levels during tumor growth in mice and humans.

Antigen stimulation in mice such as occurs with the rejection of an allogeneic tumor graft caused a substantial rise in serum glycolipid Ia levels. However, mice bearing a measurable syngeneic tumor had no detectable Ia antigens in their sera; this observation was made in several different strains of inbred mice with 5 different tumors. In each instance the serum Ia level fell as the tumor grew progressively, reached zero at about the time the tumor was visible, and remained at this zero level until the mouse died. Similar results were found in humans: Tumor-bearing patients had markedly suppressed serum Ia levels. The mechanism of the fall in serum Ia glycolipid levels is not known, but the measurement of the serum Ia glycolipid content appears to reflect the level of activation of the immune system, and the suppression of serum glycolipid Ia levels found in tumor-bearing mice and patients may have important clinical implications.

Animals↗
PubMed1980

Anti-self receptors. I. Direct detection of H-2L region-restricted receptors on murine thymocytes.

A high proportion (20--50%) of murine thymocytes form rosettes with either syngeneic or allogeneic erythrocytes. The specificity of this interaction was investigated by measuring the ability of different erythrocyte sonicates to inhibit rosette formation. With erythrocyte sonicates from recombinant mouse strains it was demonstrated that rosetting with syngeneic erythrocytes was mediated by H-2L and/or H-2D region-restricted receptors. The specificity of autorosetting was directly mapped to the H-2L region by the inability of erythrocyte sonicates from the BALB/c-H-2dm2 mutant, an H-2L-deletion mutant, to inhibit the rosetting of wild-type (BALB/c) thymocytes. The B10,2D2-H-2dm1 mutant, which has substantially modified H-2L and H-2D antigens, supported this conclusion. Furthermore, anti-H-2L sera were able to specifically block the inhibition of rosetting by erythrocyte sonicates. The above procedures clearly implicated the H-2L region in the thymocyte rosetting of d and k haplotypes. With the s haplotype the rosetting receptor was mapped to the H-2L/H-2D region, whereas with the b and q haplotypes rosetting was only mapped to the D end of the H-2 complex. This study also suggested complete cross-reaction between the thymocyte receptors carried by the k and d haplotypes, whereas the receptors of b, q, and s haplotypes were haplotype specific. In addition, the inhibition assay indicated that the rosetting of thymocytes with allogeneic and xenogeneic (rat) erythrocytes was mediated by a receptor primarily directed against self-H-2L. Finally, the critical role played by the H-2L region in this rosetting phenomenon was demonstrated by the inability of thymocytes from the H-2L-deletion mutant (H-2dm2) to rosette with syngeneic, allogeneic (rat) erythrocytes.

Animals↗
PubMed1980

Major histocompatibility gene complex (MHC)-coded determinants on antigen-specific suppressor factor for delayed-type hypersensitivity and surface phenotypes of cells producing the factor.

Antigen-specific suppressor factor for delayed-type hypersensitivity (DTH) to sheep red blood cells (SRBC) was obtained by incubating in vitro spleen cells from CBA mice (H-2k) injected intravenously 3 days previously with 1 x 10(9) SRBC. The suppressor factor was characterized for major histocompatibility gene complex (MHC)-coded antigenic determinants by passing the factor through immunosorbents coupled with appropriate alloantisera. The suppressor factor was absorbed by anti-H-2k, anti-Iak and anti-I-Jk immunosorbents but was not retained by anti-Ias, anti-I-Js, anti-I-Ak, anti-I-E/Ck or anti-H-2Kk immunosorbents. In addition, the factor bound to an immunosorbent coupled with rabbit antibodies against carbohydrate-defined Ia antigens. Furthermore, the suppressive activity that was absorbed was quantitatively recovered in the acid eluates from the immunosorbents. Treatment of the spleen cells with anti-Lyt-1.1 antiserum and complement completely abrogated their ability to elaborate the suppressor factor in vitro. In contrast, treatment with anti-Lyt-2.1 or anti-Iak antiserum and complement had no effect. Thus, it appears that the suppressor factor for DTH to SRBC bears I-J subregion-coded determinants, and its production is dependent on cells which have the Lyt-1+,2- and Ia- phenotype.

Animals↗
PubMed1980

Demonstration of carbohydrate- and protein determined Ia antigens by monoclonal antibodies.

Ten monoclonal alloantibodies were examined by submitting each antibody to five independent tests in order to determine whether they reacted primarily with the glycoprotein or glycolipid class of Ia antigens. The tests employed were as follows: (1) the ability to participate an Ia-like protein from the cell surface as detected by SDS-PAGE; (2) inhibition by protein-Ia extracts free of CHO-Ia; (3) inhibition by CHO-Ia extracts free of protein-Ia; (4) neuraminidase sensitivity of the antigen and (5) inhibition by simple sugars. Using these tests, three of the ten monoclonal antibodies were shown to recognize a CHO-Ia antigen while seven recognized the protein class of Ia antigens. The three CHO-Ia-specific monoclonal antibodies recognized Ia specificities 2, 9 and 17. Monoclonal antibodies recognizing protein-defined Ia.2 and 17 specificities were also characterized. These results imply that some Ia specificities, as defined by genetic testing, can occur both as carbohydrate-defined and protein-defined determinants.--Sugar inhibition studies showed that CHO-Ia.2 has D-glucosamine as its immunodominant sugar while CHO-Ia.17 shows preference for a beta-linked galactose. Furthermore, studies with neuraminidase demonstrated that sialic acid plays a role in the antigenic determinants of CHO-Ia.9 and CHO-Ia.17. Finally, it is noteworthy that CHO-Ia.2, the private specificity of the k haplotype, appears to be expressed only on cells and not in serum. These studies clearly demonstrate the existence of the two Ia antigen classes and emphasize the complexity of the murine I region.

Animals↗
PubMed1980

A new procedure for analysing the relationship between different cell surface antigens.

An immunoprecipitation-inhibition procedure is assessing whether different antigenic determinants are carried on the same molecule or on different molecules on cells. The procedure entails (a) exposing NP-40 lysates of cells to antibody against one antigenic specificity; (b) removing free antibody and immune complexes by absorption with protein A-bearing S. aureus bacteria; (c) adsorption of the NP-40 with a detergent binding resin; and (d) measuring the inhibitory activity of the lysates for antibody against another specificity by a rosetting assay. This method has several advantages over the widely used sequential immunoprecipitation procedure.

Animals↗
PubMed1980

The surface phenotype of a suppressor cell of delayed-type hypersensitivity in the mouse.

Delayed-type hypersensitivity (DTH) to horse red blood cells was induced in cyclophosphamide-treated CBA/H mice. The DTH reaction, represented by an increase (0 x 8--1 x 0 mm) in footpad thickness 24 h after secondary challenge, could be suppressed by the adoptive transfer of 10(7) splenic lymphocytes from syngeneic mice primed with 10(9) HRBC. The surface antigenic phenotype of the suppressor cell was determined by the formation of EA, EAC, or Ig rosettes followed by depleting the rosetted populations on Isopaque-Ficoll. The suppressor cell was found to be Ig-, FcR- and CR-, although some suppression was observed with FcR+ cells. Cell depletions with cytotoxic alloantisera and rabbit complement further characterized the suppressor cell as being Thy-1+, Ly-1+, 2-, 3-, 4-, 5+, 6+, 7-, Ia- and IJ-. This cell surface phenotype if unique and differs from the Ly-1-, 2+, 3+, I-J+ suppressor cell of antibody formation and from the recently described Ly-+, 2+, 3+ feedback suppressor T cell.

Animals↗