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D G Braun

Publications and source records attributed to D G Braun.

At least 73 records · Page 4Linked to original sources
PubMed1979

Distinct functions of monoclonal IgG antibody depend on antigen-site specificities.

Intraveneous hyperimmunization of selectivity bred rabbits with streptococcal group A-variant vaccines elicits antibody responses of restricted heterogeneity at high antibody levels. All antisera contain two functionally distinct antibody populations, which can be isolated in single-band purity upon analytical isoelectric focusing. Typical examples of these two kinds of single-band antibodies were investigated in great detail for several parameters by a variety of methods. 85--99% of the streptococcal group A-variant polysaccharide (Av-CHO)-specific antibody in the antisera does not precipitate the isolated 5,000 daltons poly-L-rhamnose antigen, neither agglutinates nor lyses in the presence of complement Av-CHO-coated sheep erythrocytes (SRBC), binds the radio-labeled Av-CHO with an association constant in the ragne of 10(5)--10(6) M-1, and is of terminal specificity (nonreducing end) for the linear Av-CHO. In contrast, the minor fraction of Av-CHO-specific antibody (1--15%) does precipitate the linear Av-CHO, both agglutinates and lyses Av-CHO-coated SRBC in the presence of complement, has an affinity range of 10(8)--10(9) M-1, and is of internal specificity for the Av-CHO. The antigenic determinants of the Av-CHO for the antibodies are nonoverlapping, only one Fab of the low affinity antibody can be bound whereas four Fab of the high affinity antibody are accommodated. Hence, the determinant specificity explains the functional differences observed, for there is no indication of subclass differences. A mechanistic model of the A-variant carbohydrate presentation on the vaccine appears to account best for the unbalanced levels of low and high affinity antibody.

Animals↗
PubMed1979

Clonal dominance of low-affinity antibodies in rabbit hyperimmune anti-streptococcal group A-variant polysaccharide antisera.

Intraveneous hyperimmunization of selectively bred rabbits with streptococcal group A-variant vaccines elicits antibody responses of restricted heterogeneity at high antibody levels. In these antisera, IgG with dissociation constants Kd = 10(-6) M constitutes 90% and IgG with Kp = 10(-9) M accounts for only 10% of the group A-variant polysaccharide-specific antibodies. The low affinity antibody fraction represents the dominant clonotypes. Preparative isoelectric focusing in granulated (Ultrodex G-75) gels was used to successfully purify single-band material belonging to dominant spectrotypes. Affinity studies with these antibody fractions with the highest reported degree of purity yielded Kd = 10(-6) M values, thus confirming that clonal dominance is exclusively associated with low-affinity antibodies. Since it is known from previous work (M. Cramer and D. G. Braun, Scand. J. Immunol. 1975. 4:63) and from the rabbit antisera used here that clonal dominance of this sort is long-lived, this work fails to support the argument of immune maturation. The data more logically relate to antibodies that emerge with different subspecificities -- recognized in the antigen as a function of time in immunization procedures -- rather than to an inherent property of the immune system.

Animals↗
PubMed1979

Rabbit allotype a locus subspecificities of homogeneous anti-streptococcal antibodies.

Rabbits were immunized with group A-variant (Av) streptococcal vaccine to elicit antisera containing antibodies of "restricted heterogeneity" specific for Av polysaccharide. Antibodies that migrated as single bands in polyacrylamide gel isoelectric focusing were isolated on agarose block electrophoresis and preparative isoelectric focusing. These homogeneous antibodies were analyzed for content of the a locus markers by inhibition of allotype-anti-allotype radioimmunoassays. Both single-band preparations and preparations containing 2--5 bands were deficient in content of the a locus markers. These analyses were strongly influenced by the different anti-allotype antisera used. Single-band preparations, deficient in a 2 or a 3 markers, were further used to produce anti-allotype antisera, and these sera were assayed for reaction with pooled a2 or a3 molecules. The results, in contrast to the a1 allotype, failed to demonstrate common determinants on either a 2 or a 3 molecules indicating that each a allotype consists of a set of different Ig molecules. The data suggest different evolutionary pathways of the a 1, as opposed to the a 2 and a 3 molecules.

Alleles↗
PubMed1979

The VkVI subgroup of rabbit light chains: complete amino acid sequence of a third variable region (K29-213).

The amino acid sequence of the variable region of a rabbit anti-streptococcal A-variant antibody light chain was determined. By using a combination of different cleavage methods, the sequence was established. Large peptides were sequenced in an extensively modified Beckman sequentor. Light chain K29-213 belongs to a rare subgroup (kVI). Several of these light chains of antibodies with different specificities have been totally or partially sequenced. Comparison of these light chains reveals at least four germ-line-encoded variants within this subgroup.

Amino Acid Sequence↗
PubMed1979

Clonotype patterns of antibodies released by single lymph nodes.

Experimental primary, secondary and tertiary stimulation with streptococcal vaccine of isolated lymph nodes in vivo in the sheep model induces largely persistence of anti-group polysaccharide antibody clonotype patterns with the rare occurrence of additional clonotypes demonstrable after secondary stimulation persisting during the tertiary stimulus. It is not clear whether these additional clonotypes are the products of mutants or whether they pre-existed and were demonstrable only after a secondary stimulus, because of threshold concentrations required for identification. Further, isolated contralateral popliteal and prescapular lymph nodes of individual sheep share completely overlapping clonotype patterns during experimental primary anti-polysaccharide antibody responses indicating an identical repertoire of specific clonotypes under this condition of responsiveness.

Animals↗
PubMed1979

T cell activation of primed rabbit blood lymphocytes by antigen.

Activation of primed rabbit PBL by homologous antigen in the early proliferative phase (on days 3--5) mainly involves lymphocytes which neither secrete specific antibody nor contain immunoglobulin in their cell membrane. This stimulation is antigen-specific, and evidence is given that the major proportion of these cells are T lymphocytes. The B cells forming Av-CHO-specific PFC were studied by autoradiography on days 6 and 12 of culture. Since incorporation of radioactive thymidine was found in the majority of PFC, these cells are also in proliferation.

Animals↗
PubMed1979

Persistent clonotypes associated with group A streptococcal polysaccharide antibody response in man.

Elevated anti-streptococcal group A polysaccharide antibodies are present in the sera of acute rheumatic fever patients. Analytical isoelectric focusing of sera taken in the acute phase of the disease and sequentially for up to 2 years later reveals a stable clonotype pattern of anti-group A polysaccharide antibodies. In addition, with reinfection and recurrence of the rheumatic fever, the same clonotype pattern of antibodies is present. This finding indicates that naturally acquired immunity is associated with persistence of clonotype expression.

Antibodies, Bacterial↗
PubMed1979

Amino acid sequences of antibody light chain variable regions of pedigreed rabbits: kappa light chain K49-501 (allotype b4 anti-streptococcal group A-variant polysaccharide antibody).

The amino acid sequence of positions 1--150 of a light chain, isolated from another monoclonal rabbit anti-streptococcal group A-variant polysaccharide antibody, was determined. The analysis was performed with 2 mumol of polypeptide chain, using a grossly modified Beckman 890B sequenator. This sequence stretch accounts for the whole variable region and a considerable part of the constant region at a total length of 218 amino acids. This allotype b4 light chain was isolated from a non-precipitating, end-group-specific antibody with a KD = 1.3 X 10(-5)M. This brings the present number of totally known rabbit VL sequences of antigen elicited antibodies to 21. A comparison of these 21 sequences reveals a building plan of ribbit VL homologous to that of human and murine VL regions. The observed variability does follow a pattern of linked amino acid substitutions, indicating that this information must be contained in the germ-line of the rabbit in the form of multiple VL region genes. This conclusion, however, does not rule out the occasional variant being due to somatic rearrangement. Finally, this comparison reveals that the joining peptide between positions 96--110 is also a separate entity in rabbit VL region sequences.

Amino Acid Sequence↗
PubMed1979

Light chain heterogeneity of type lambda anti-streptococcal group A-variant polysaccharide antibodies in rabbits.

BASILEA rabbits lack the expression of chi-polypeptide chains and compensate for this lack with expression of lambda polypeptide light chains. These rabbits were immunized with streptococcal group A-variant vaccines. The heterogeneity of the lambda polypeptide chains of specific antibodies was analyzed and compared with that of chi light chains. No significant difference was found by SDS-PAGE and IEF in the number of light chain bands of high affinity antibody expressed. This suggests that in the rabbit the size of the variable region repertoire is similar for lambda and chi light chains.

Animals↗
PubMed1978

Activation of primed rabbit blood lymphocytes by antigen: early phase of triggering and the specificity of the response.

The early phase of antigen-dependent triggering of rabbit blood lymphocytes (PBL) is described. Specific activation of PBL by streptococcal vaccines requires primed lymphocytes from high responder rabbits. B-cell memory is conferred by Ig receptors by both mu and gamma chains. Antigen-induced incorporation of leucine and thymidine reach peak values at day 5, with stimulation indices of 10 to greater than 100, closely followed by increases in the number of living cells. A second and less intensive phase of cell proliferation is seen on days 8--11. A large fraction of cells (between 3 and 33%) participates in proliferation. A rough calculation of the average doubling time of cells during the exponential growth phase gives values of 8--16 h.

Animals↗
PubMed1978

Variable-region subgroup and specificity of cold agglutinins.

The variable-region subgroup determined by amino acid sequence analysis of heavy and light chains of two monoclonal cold agglutinins with the new anti-Gd specificity is reported. Both proteins belong to the VHIII subgroup of heavy chains; one light chain falls into the V kappaI subgroup, the other has a blocked N-terminus which so far has not been observed in human kappa chains. The comparison of anti-Gd with anti-I/-i or anti-Pr cold agglutinins indicates that anti-Gd differs from other cold agglutinins with respect to variable-region subgroup. The data extend previous findings on the restriction of certain antibodies to distinct variable-region subgroups.

Agglutinins↗
PubMed1978

Specific and cross-reactive activation of rabbit peripheral blood lymphocytes by streptococcal vaccines.

The ability of primed rabbit blood lymphocytes to respond in vitro to the homologous streptococcal group antigen depends on its presence in culture in the form of vaccine or cell walls thereof. These lymphocytes can also be stimulated in vitro by streptococcal vaccines with chemically related group antigens. The basis for this cross-stimulation apparently resides in shared rhamnose moieties. Activation of these lymphocytes was not achieved by vaccines from unrelated bacteria. There is also the suggestion that rabbits of different genetic origin differentiate between cross-stimulating antigens, probably at the level of antigenic determinants. The data support the view that recognition and response pattern of the immune system rely heavily on a network of antigens.

Animals↗
PubMed1978

Rabbit variable kappa light chain regions: subgroups contain polypeptides encoded by multiple genes.

Two identical light chain variable regions were identified in anti-streptococcal Group A-variant antibodies elicited in litter-mate rabbits by hyperimmunization with vaccine. In addition, one rabbit produced two additional clonally restricted antibodies to this polysaccharide antigen. The partial amino acid sequence of the light chain of one of these antibodies was identical with the dominant antibody light chain sequence, while the light chain of the other antibody, also partially established, showed significant variations in the framework-associated regions with identical CDRI and II. Since all of these light chains were from a small subset of rabbit kappa light chain pools (b4 allotype) the data suggest, together with other light chains reported in the literature, that more than one copy of variable region genes are present in the germ-line per subgroup. Furthermore, framework associated amino acid substitutions are not random; this suggests the existence of some "ordered" mechanism for linked amino acid substitutions (presumably recombination). Furthermore one light chain can pair with more than one heavy chain to yield functional antibodies.

Amino Acid Sequence↗
PubMed1977

Antibody response to the streptococcal group A-variant polysaccharide in BASILEA rabbits lacking kappa-polypeptide chains.

Rabbits from a variant strain called BASILEA, in which homozygotes express only lambda-type chains and heterozygotes have normal kappa/lambda ratios, were hyperimmunized with a streptococcal group A variant vaccine. Homozygotes (bas/bas) produced antibodies with lambda-chains, heterozygotes, however, produced predominantly antibodies with kappa-chains. The incidence of restricted high responders in the BASILEA strain was high; it was probably introduced by the original mutant rather than by the loss of kappa-chains (founder effect). The degree of heterogeneity of homozygotes is similar to the heterogeneity of heterozygotes, and to that of rabbits expressing kappa-chains. This suggests that in the rabbit, the repertoire of lambda-chain genes is of similar size to that of kappa-chain genes.

Animals↗
PubMed1976

Antipolysaccharide antibodies of restricted heterogeneity secreted by a single lymph node.

Immune responses which give rise to the synthesis of antibodies of restricted heterogeneity can be reproducibly induced in rabbits and mice by streptococcal polysaccharide antigens (1,2). While these reports have demonstrated clonotype restriction in the immune serum of rabbits and mice, they give little information with respect to the clontype restriction in a single reaction site of organized lymphoid tissue, for example a single lymph node. The kinetics of immune responses in lymph nodes in situ have been studied in sheep using a variety of antigens (3-6), and a substantial body of information is available on the changes in cell output, cell type, and the number of antibody-secreting cells which occur within the efferent lymph of antigen-stimulated lymph nodes. However, there is no information with respect to the amount and the heterogeneity of antibody that is secreted by the lymph node or cells within the efferent lymph. In the present report we have examined the temporal sequence of clonal restriction in the efferent lymph of individual sheep popliteal lymph nodes undergoing an immune response to the streptococcal group A-variant polysaccharide (Av-CHO).

Animals↗