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Biomedical subjects

G B Ferrara

Publications and source records attributed to G B Ferrara.

At least 127 records · Page 7Linked to original sources

Serologically detectable polymorphism of the HLA-DC alpha-subunit.

Two 8th Workshop alloantisera, 8W1062 and 8W614, which were classified respectively as anti-DR5 and anti-DR3, were found to react with an Ia preparation from LG38 cells (DR5,5) that was depleted of DR and BR molecules and enriched in DC molecules by pretreatment with a monoclonal antibody and an antiserum against alpha-subunits of DR and BR molecules. The reaction of both alloantisera was inhibited by DR3-positive, DR5-positive, and DRW13-positive individuals. This pattern does not correspond to any of the hitherto reported supertypic specificities. Both antisera were shown to bind to the alpha-subunit isolated from the DC-enriched preparation but not to the beta-subunit. The specificity has been named DC alpha 3. Family studies show the segregation according to the HLA haplotype patterns. This demonstrates by formal genetics criteria the HLA linkage of the locus controlling the DC alpha-subunit.

Binding, Competitive↗

Three Ia species with different structures and alloantigenic determinants in an HLA-homozygous cell line.

Three distinct molecular subsets with different structures and alloantigenic determinants were identified in human Ia antigens from cells of an HLA-Dw7 homozygous cell line. The subsets carried DR7 specificity, BR4X7 supertypic specificity and MB2 supertypic specificity, respectively, and were immunospecifically separated by the use of operationally monospecific alloantisera. These specificities showed HLA-linked segregation in families and they were distributed in the population according to different but partially overlapping patterns. On peptide mapping analysis, the three subsets showed marked differences in the beta-chains. The alpha-chains of DR7 and BR4X7 subsets were very similar to each other, whereas the alpha-chains of MB2 subset were distinctive from those of DR7 and BR4X7. These data indicate the presence of a minimum of three HLA-linked loci; DR locus, a locus that encodes BR4X7, and a locus that encodes MB2, and substantiate the three-loci concept for the genetic control of human I antigens.

Amino Acid Sequence↗

The distribution of DR5, MT2, and MB3 specificities on human Ia subsets.

Human Ia molecules were isolated from cells of LG-38, an HLA-homozygous lymphoid cell line of DR5 specificity. Three Ia subsets could be distinguished and separated by using specific alloantisera and a monoclonal antibody with polymorphic reactivity. These subsets carried the specificities DR5 and MT2, MT2 alone and MB3 alone. The structure of the three molecular species was analyzed by microfingerprinting. The subset carrying only MT2 was similar, in both the component alpha and beta chains, to the major subset carrying DR5 and MT2, whereas the subset carrying MB3 was distinct in both chains from the other two subsets. These data are compatible with our previous findings obtained for the products of two Ia loci closely linked to the DR locus and provisionally called DC and BR; they also support the conclusion that the subset carrying only MT2 is an allelic product of the BR locus, whereas the MB3 subset is an allelic product of the DC locus. MT2 appears to be a shared specificity of DR and BR loci products.

Antibodies, Monoclonal↗

Evidence that celiac disease is primarily associated with a DC locus allelic specificity.

Sixty patients with celiac disease were typed by radioimmunoassay for the specificities HLA-DR3, HLA-DR7, and for an allelic specificity, DC3, of the HLA-associated DC locus. We found that celiac disease is primarily associated with the DC determinant. The previously described associations with DR3, DR7, B8, B13, and A1 can be explained by decreasing degrees of linkage disequilibrium with DC3.

Adolescent↗

Antigen-specific antibody response induced in cultures of human blood lymphocytes in presence of polyethylene glycol.

Human peripheral blood lymphocytes respond in vitro to sheep red blood cells (SRBC) with the appearance of numerous specific plaque-forming cells, if the Epstein-Barr virus (EBV) is added to the cultures together with the antigen. However, it was found that when polyethylene glycol (mol. wt. 6000) is included in the culture medium at a final concentration of 4%, antigen alone is able to induce an anti-SRBC response, with clear hemolytic plaques of regular size. Addition of EBV causes a further increase in the antigen-specific antibody response.

Animals↗

HLA segregation of human T-cell subpopulation markers.

Antisera recognizing subsets of human T lymphocytes can be produced by planned immunizations involving HLA-A and HLA-B compatible donors. The reactivity of these antisera against some individuals of a population, but not others, show that they recognize a polymorphic cell surface component. The study of informative families shows the HLA linkage of at least some of these alloantigenic determinants.

Antibody Specificity↗

Human T lymphocytes subpopulation as defined by alloantigens and FC receptors a comparative analysis.

Human peripheral T cells were fractionated in accordance to their surface receptors for G or M immunoglobulins and analyzed for their reactivity with alloantisera obtained by planned immunization involving HLA-A and HLA-B compatible individuals. These alloantisera were previously shown to recognized polymorphic structure exclusively expressed on T cells. Although the majority of the alloantisera analyzed reacted with different proportions of both T(M) and T(G) populations, some antisera specifically recognized surface structure restricted to either T(M) or T(G) lymphocytes. The various alloantisera consistently reacted with a fraction only of T(G) and T(M) cells, this indicating that these T cell subsets can be further fractionated in accordance to the expression of the alloantigenic determinants recognized by these antisera.

Animals↗

Further characterization of two "DR supertypic" specificities. Additional evidence that they reside on molecules different from those carrying HLA-DR locus specificities.

Peripheral lymphocytes from a panel of individuals who had been assayed for DR specificities by the conventional cytotoxicity assay were typed for DR "supertypic" specificities, DC1 and BR4 x 7, by the radioimmunoassay. A positive and a negative population were clearly distinguished for both specificities and the strong association of the DC1 specificity with DR1, 2, and w6 was confirmed as well as the BR4 x 7 specificity with DR4 and 7. Family study also supported this strong association. Appropriate papain digestion separated molecules carrying DC1 determinant from those carrying DR2 as well as from those carrying DRw6, and separated molecules carrying BR4 x 7 from those carrying DR4. Specificity analysis of the 8th Workshop antisera by use of these separated antigen preparations showed that some anti-DC1 antisera do not possess appreciable anti-DR1, 2, or w6 activity and vice versa. The same was found for DR4 x 7 in its relationship with DR4 and 7. The existing evidence could be explained most economically by assuming a genetic model of two loci in linkage disequilibrium each coding for analogous but distinct forms of the small (beta) subunits of Ia molecules.

Epitopes↗

Human B-cell alloantigens DC1, MT1, and LB12 are identical to each other but distinct from the HLA-DR antigen.

Some human B-lymphoblastoid cell lines are shown to express at least two types of Ia-like antigens. One antigen is defined by alloantisera to HLA-DR, and the other antigen is defined by alloantisera and a monoclonal antibody to the specificities DC1, MT1 (MB1), and LB12, which are in linkage disequilibrium with HLA-DR. The subunits of the DC1 molecule differ from those of the DR molecule. The light chains of both molecules are structurally polymorphic.

B-Lymphocytes↗

Immunochemical definition of the new dr specificity 8WDRw13.

Using two alloantisera produced by planned immunization of blood donor volunteers, a new specificity was defined residing on human Ia molecules. The specificity is possessed by the following human lymphoid cell lines: Reh, NALM-1, B89, KM-3 and BJAB, and appears to be the same as the DRw13 specificity discovered in the 8th International Histocompatibility Workshop. A radioimmunoassay for typing w13 on peripheral blood lymphocytes was established. A population and family study supports the genetic control by the HLA-DR locus. The phenotypic frequency in an Italian population was 3.7 percent. The relationship between w13 and the MT2 supertypic specificity was confirmed at the molecular level and the papain sensitivity of w13-carrying molecules was determined.

Cell Line↗

Molecular identification of human Ia antigens coded for by a gene locus closely linked to HLA-DR locus.

Human Ia(-like) specificities controlled by gene loci other than HLA-DR were searched for at the molecular level in cells of human B-cell-type cell lines which carry two established DR specificities. Chevalier cells of DRw3 and 7 and U698M cells of DRw2 and 4 were used. Their Ia molecules were partially purified, radioiodinated and analyzed for Ia specificities by the direct binding and sequential binding assays with a selected panel of human Ia alloantisera. It was possible in both the cell lines to define a third subset of Ia molecules carrying a new specificity in addition to two Ia subsets carrying the established DR specificities. The new specificity was detected by putative anti-DRw4 and anti-DRw7 antisera and was closely associated with DRw4 and DRw7 at population level. It was thus designated provisionally as BR4X7. These results suggest that the BR4X7 specificity is coded for by a separate Ia locus closely linked to HLA-DR locus. The determinant(s) responsible for BR4X7 was located on the small subunit of Ia molecules.

Animals↗

HLA-DR typing by radioimmunoassay.

A radioimmunoassay procedure is described by which peripheral blood lymphocytes can be typed for HLA-DR specificities. The major advantages of this method are the following: simple and reproducible procedure, no need for B lymphocyte separation, no need for optimal viability, and no need for preabsorption of antisera with platelets. This method will find an application in the genetic and biochemical analysis of the HLA complex, and in the clinical tests of Ia antigens for diagnostic or prognostic purposes and in retrospective transplant studies.

Antibody Specificity↗

The HLA system in the Campania region: a genetic study.

The HLA-A, -B and -C locus gene frequencies and the significant A, B and B, C haplotypes of the population of Campania (Southern Italy) are reported. From a comparison of gene frequencies and from an overall genetic distance evaluation with other European and Mediterranean populations the typical Mediterranean structure of the Campanian population is confirmed. While keeping several Middle Eastern features, it places itself as far from Northern Italy (Bergamo) as from the Spanish and the French populations. Among all the groups compared, including Turkish and Lebanese, the largest distance is measured from the English, the Danish and the German populations.

Chromosome Mapping↗

Markers of human T cell subsets identified by alloantisera.

Analysis by the indirect fluorescence test followed by fluorescence-activated cell sorter (FACS) analysis has shown that antisera recognizing subsets of human T lymphocytes can be produced by planned immunizations involving HLA-A and HLA-B compatible donors. The reactivity of these antisera against some individuals of a population but not others shows that they recognize a polymorphic cell surface component. The reactive subpopulation largely overlaps with the JRA+ subset, which was previously shown to possess regulatory properties in functional assays. The specificity of the antisera for a T cell subset and the unrelatedness of the anti-B cell activity of the same antisera has been confirmed by two-color fluorescence tests.

Antibody Specificity↗