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Biomedical subjects

M Potter

Publications and source records attributed to M Potter.

At least 235 records · Page 13Linked to original sources
PubMed1983

Galactan-binding antibodies. Diversity and structure of idiotypes.

A group of eight IgM hybridoma proteins induced with beta(1,6)-D-galactan-containing antigens has been characterized in terms of primary amino acid sequence and idiotype expression. The H chain amino acid sequences reveal very strong homology in the VH segment although several substitutions are seen that suggest the occurrence of somatic mutation in these IgM molecules. Significant sequence variation was observed in CDR-3, the region generated by the D segment, and the two recombination events, VH-D and D-JH. The number of amino acids in this region contributed by the D segment was found to vary from two to six, yet the overall length of CDR-3 was precisely maintained by the addition of amino acids on either side of D during the recombination processes. These additional amino acids are suggested to result from nucleotide addition by repair enzymes. Idiotypic analysis of these proteins, in conjunction with an assessment of the H chain sequences, has permitted an identification of the molecular basis of both cross-reacting and unique idiotypic determinants expressed by these molecules.

Amino Acid Sequence↗
PubMed1983

DNA rearrangement and altered RNA expression of the c-myb oncogene in mouse plasmacytoid lymphosarcomas.

Three types of tumors termed plasmacytomas (ABPC's), lymphosarcomas (ABLS's), and plasmacytoid lymphosarcomas (ABPL's) arise in BALB/c mice treated with pristane and Abelson murine leukemia virus (A-MuLV). While most ABPC's and BLS's contain integrated A-MuLV proviral genome and synthesize the v-abl RNA, most ABPL's do not. The ABPL tumors were examined for the expression of other oncogenes that may be associated with their transformed state, in the absence of transforming virus. These tumors expressed abundant c-myb RNA of unusually large size and showed DNA rearrangements of the c-myb locus.

Abelson murine leukemia virus↗
PubMed1983

Polymorphism in anti-phosphocholine antibodies reflecting evolution of immunoglobulin families.

Complete variable (V) region amino acid sequences were determined for four heavy (H) and one light (L) chain from C57BL phosphocholine (PC)-binding monoclonal antibodies. Additional NH2-terminal sequences were obtained from H and L chains of C57BL and CBA/J origin. When these V regions were compared with previously reported anti-PC sequences, a number of observations could be made regarding the function and evolution of L and H chain segments used in these antibodies. (a) L and H chain V segments are remarkably conserved in these inbred strains, although there has been an accumulation of point mutations identifying apparently allelic forms of VK and VH. (b) Mice of each genotype use the same three VK segments in combination with a single VH segment to produce most anti-PC antibodies. An exception has been noted that indicates the occasional use of a second VH gene segment. (c) Multiple, different DH regions are used by mice of each strain, which suggests that the DH segment sequence plays no critical role in either antigen binding or VH-VL pairing. Furthermore, the DH segments and their corresponding gene families appear to be highly conserved in the inbred strains studied. (d) Most PC-binding antibodies use the JH1 joining segment. All JH1 sequences from C57BL mice differ from the BALB/c JH1 at position 105, which identifies allelic forms of the JH1 region. These studies are a first assessment of the nature of mutational events associated with the evolution of specific multigene immunoglobulin families and indicate that homologous VH, DH, JH, VK, and JK genes are similarly assembled and expressed in PC antibodies from three diverse genotypes.

Amino Acid Sequence↗
PubMed1983

Structural correlates of immunoglobulin diversity.

VL and VH domains, from different species and with widely different primary structures, interact with each other in the same way to create the globular FV region. Much of the FV is a highly conserved framework structure that is probably common to most, if not all, mammalian FV regions. The extensive contoured frontal surface of the FV is composed of highly variable polypeptide segments (Wu-Kabat complementarity-determining regions). These segments are derived from parts of VL, VH, JH gene products and most of the D gene product. This surface is currently considered to be the most likely location of the antigen-binding sites. The firm immunochemical data based on identification of contacting amino acids supporting this location are still, however, very fragmentary. VL and VH gene products form a large part of the potential antigen-reactive surface. Hence, combinations of different VL and VH gene products are the largest source of structural diversity. The JL and JH gene products have chiefly structural functions in maintaining domain architecture and controlling some interactions between VL and VH domains. The VL-J junction amino acid can provide unique structural properties in the deeper accesses of the potential antigen reaction surface. The VHD-JH junction is more superficial and could be, but has not yet been, directly implicated in antigen binding. The D gene product and the additional amino acids associated with the (VH-D-JH) rearrangement process do determine a substructural part of the potential antigen reactive surface. The D gene product (a connecting segment between two beta strands) can have many different secondary structures. Functionally, the D region product could interact with VL-CDR-1 amino acids to create a specific contour of the antigen reaction surface. Curiously, primary structural variations in H3 have not yet been directly implicated in antigen binding. Much remains to be learned about the role of VH-D-JH rearrangement in antibody diversity. The major genetic factors in creating structural diversity are the multiple VL and VH gene libraries. The gene rearrangement process provides a further amplification. Somatic mutations are yet another additional mechanism.

Animals↗
PubMed1983

Increased expression of myc-related oncogene mRNA characterizes most BALB/c plasmacytomas induced by pristane or Abelson murine leukemia virus.

RNA blots of poly(A)-containing RNA from normal livers and spleens and from a number of transplantable hematopoietic and lymphoid BALB/c tumors, including early and late generation plasmacytomas, were hybridized with probes for four onc genes. abl RNA was abundant only in those tumors producing Abelson virus, bas RNA was found in approximately equal amounts in normal tissues and plasmacytomas, and myb RNA was absent in normal liver and plasmacytomas. Normal liver and spleen RNA showed faint traces of myc hybridization, but myc RNA was increased in most plasmacytomas. In one plasmacytoma, TEPC 1165, a particularly abundant amount of myc RNA was found, principally as a 3.5-kilobase band. In the other plasmacytomas, bands of 2.4- or 1.8-kilobase myc RNA were found. Southern blots of DNA from tumors that contained 2.4-kilobase or larger myc RNA showed myc hybridization to an EcoRI fragment of about 21 kilobase pairs, similar to the myc band in normal DNA. EcoRI digests of DNA from two tumors that expressed myc RNA of 1.8 kilobases showed an additional smaller myc band, suggesting that the myc gene is rearranged in these plasmacytomas. The basis for increased myc gene transcription in plasmacytomas is not understood, but the evidence suggests that different mechanisms may be operating in different plasmacytomas. Apparently, neither myc gene amplification nor myc gene rearrangement is required for increased myc transcription.

Abelson murine leukemia virus↗
PubMed1983

A BALB/c congenic strain of mice that carries a genetic locus (Ityr) controlling resistance to intracellular parasites.

BALB/c.DBA/2 Idh-1b-Ityr-Pep-3b congenic mice were developed by introgressively backcrossing the Idh-1b and Pep-3b markers of DBA/2 mice onto the BALB/c pi mice. This introduced a 30-centimorgan chromosome 1 segment of DBA/2 chromatin that contained the Ityr gene. BALB/c.DBA/2 Idh-1b-Ityr-Pep-3b mice were resistant to in vivo infections by Salmonella typhimurium, Mycobacterium bovis, and Leishmania donovani.

Animals↗
PubMed1983

Peritoneal plasmacytomagenesis in mice: comparison of different pristane dose regimens.

Plasmacytomas were induced in inbred BALB/c pi mice by the ip injection of pristane with 4 different dose schedules. Three 0.5-ml doses (1.5 ml) given at 2-month intervals gave an average yield of 61% plasmacytomas in 6 experimental groups with a range from 51 to 71%; a single 1-ml dose gave an average yield of 42% plasmacytomas in 5 experimental groups with a range from 37 to 45%; and a single 0.5-ml dose gave an average of 22% from 3 experiments involving young mice with a range from 14 to 26%. Two 0.5-ml doses given at various intervals from 14 to 300 days gave yields of plasmacytomas that usually but not always were greater than that obtained with a single 0.5-ml dose of pristane. When the second injection of pristane was delayed as long as 180 days, a strong additive effect over that observed with 0.5 ml alone was obtained. The plasmacytomas developed in mice given the second dose 180 days after the first, with virtually the same latent period as observed with a single 1-ml dose. No plasmacytomas were found in 200 BALB/c pi mice inoculated with corn oil, aluminum hydroxide, or very small doses of pristane (i.e., 0.05 ml). The minimal latent period for plasmacytoma development is about 120 days. The median latent period ranged from 180 to 250 days in the groups of mice that received 3 0.5-ml injections of pristane. In a single experiment pristane freed of UV-absorbing materials was as potent as commercial grade pristane in inducing plasmacytomas.

Animals↗
PubMed1983

An allotype linked gene that is associated with a negative or very low anti-phosphorylcholine response (PC) phenotype in wild mice (CNV).

In contrast to most inbred and wild mice, a population of wild mice recently isolated from a farm in Centreville, MD, and designated CNV produced no anti-phosphorylcholine (PC) antibodies (less than 1 microgram/ml) in response to immunization with the PC antigen Streptococcus pneumoniae (R36A) and gave 9 to 36 micrograms/ml anti-PC response to PC-KLH at 14 days after immunization. When another carbohydrate antigen, namely, bacterial levan, was used, CNV mice all gave high antibody titers. When CNV (PC-) mice were bred to inbred C.B20 (PC+) mice, 82% of the F1 and 76% of the F2 hybrids were surprisingly non-responders (PC-), which suggested that PC- gene(s) of CNV origin dominated the response to these antigens. The 18% PC+ phenotype in the F1 hybrids indicated possible heterozygosity of the PC genes controlling the PC- response in the CNV mice. Genetic studies on CNV mouse No. 378 supported this possibility. Analysis of the F2 data strongly suggest that two genes determined the PC- response, one of which was closely linked to the Igh-C allotype locus (chromosome 12). Hypothetically, we propose that CNV mice have two genes that cooperate but that sometimes act independently to express the PC- phenotype. Surprisingly, when F1 mice giving PC- phenotypes were back-crossed to C.B20, very few mice (18%) were PC-. This indicated that the PC- determining genes of CNV origin were not able to dominate immune responses in the presence of a larger number of C.B20 genes. This kind of expression may be regulated by other factors, such as clonotype competition and clonal dominance.

Animals↗
PubMed1982

Amplification of immunoglobulin lambda constant genes in populations of wild mice.

The lambda immunoglobulin light chain (Ig lambda) locus of BALB/c inbred mice consists of two variable region gene segments (V lambda)1-3, and four constant region gene segments (C lambda)1,2,4,5. Each C lambda gene segment is associated with a unique joining segment (J lambda)2,4-7, and they are organized in two paired units, J3C3-J1C1 and J2C2-J4C4 (refs 4, 8). Using cDNA probes specific for C lambda 1 and C lambda 2 (ref. 9) we have analysed the genomic organization of the C lambda gene segments in wild-derived and inbred strains of mice. Although Southern blots of the genomic DNA of inbred mice show a constant pattern of hybridization, wild-derived mice show a high degree of variation in the number, size and intensity of hybridizing fragments. We have now found that, per haploid genome, mice of a Mus musculus musculus stock isolated from Sladeckovce, Czechoslovakia (CzII) have at least 12 C lambda segments, and mice of a Mus musculus domesticus stock 'Centreville Lights' from Centreville, Maryland (CL) have at least 8 C lambda segments. There appears to have been relatively recent amplifications of the C lambda gene segments in wild mice.

Animals↗
PubMed1982

Classification of mouse VK groups based on the partial amino acid sequence to the first invariant tryptophan: impact of 14 new sequences from IgG myeloma proteins.

Fourteen new VK sequences derived from BALB/c IgG myeloma proteins were determined to the first invariant tryptophan (Trp 35). These partial sequences were compared with 65 other published VK sequences using a computer program. The 79 sequences were organized according to the length of the sequence from the amino terminus to the first invariant tryptophan (Trp 35), into seven groups (33, 34, 35, 36, 39, 40 and 41aa). A distance matrix of all 79 sequences was then computed, i.e. the number of amino acid substitutions necessary to convert one sequence to another was determined. From these data a dendrogram was constructed. Most of the VK sequences fell into clusters or closely related groups. The definition of a sequence group is arbitrary but facilitates the classification of VK proteins. We used 12 substitutions as the basis for defining a sequence group based on the known number of substitutions that are found in the VK21 proteins. By this criterion there were 18 groups in the Trp 35 dendrogram. Twelve of the 14 new sequences fell into one of these sequence groups; two formed new sequence groups. Collective amino acid sequencing is still encountering new VK structures indicating more sequences will be required to attain an accurate estimate of the total number of VK groups. Updated dendrograms can be quickly generated to include newly generated sequences.

Amino Acid Sequence↗
PubMed1982

The interaction of mouse myeloma immunoglobulin S15 with negatively charged polysaccharide antigens.

A murine BALB/c IgG2a (lambda 3) myeloma immunoglobulin SAPC-15 with binding activity for negatively charged polysaccharides has been purified by affinity chromatography, and its interaction with heparin and various other polyanionic antigens has been studied. The antigen-binding activity has been demonstrated to reside in the Fab part of the immunoglobulin. The S15 myeloma protein in 0.05 M Tris buffer at pH 7.4 precipitated dextran sulfate, heparin, chondroitin sulfate A, B and C, hyaluronic acid, H. influenzae type b polysaccharide, calf thymus DNA, Klebsiella polysaccharide K63 and poly-L glutamic acid. Of these antigens only dextran sulfate was precipitated in 0.01 M phosphate buffered saline (0.15M), pH 7.4. The pepsin S15 Fab fragment did not precipitate with any of these antigens. The intrinsic tryptophanyl fluorescence of S15 was changed maximally by the addition of heparin, and the binding affinity of the immunoglobulin for this antigen was high (greater than 10(6) L/M). S15 may resemble antibody molecules that react with antigens under non-physiological conditions or in pathological conditions or in the external environment as in the lumen of the gut. All the above interactions of S15 with antigens persisted in 0.05 M Tris buffer made physiologically isotonic by the addition of sucrose, and S15 could thus be used to identify these antigens on cell surfaces.

Animals↗
PubMed1982

Novel class of mouse mammary tumor virus-related DNA sequences found in all species of Mus, including mice lacking the virus proviral genome.

Mice in breeding colonies of feral Mus musculus brevirostris (Azrou, Morocco), M. m. musculus (Studenec, Czechoslovakia), and M. m. molossinus (Fukuoka, Japan) were found to lack the mouse mammary tumor virus (MMTV-alpha) proviral genome in their germ line. MMTV-alpha proviral genomes have been found in all inbred strains of M. musculus by using high-stringency nucleic acid hybridization conditions. We conclude that feral mice in these colonies are heterozygous for a limited number of MMTV-alpha proviral genomes and that those lacking them arose as a result of random chromosomal segregation. All mice in another breeding colony of feral M. m. musculus (Sladeckovce, Czechoslovakia) lack MMTV proviral genes. By relaxing the conditions of nucleic acid hybridization, MMTV-related sequences (designated MMTV-beta) were detected in restricted cellular DNA from MMTV-negative mice and all other inbred strains and feral species of the genus Mus. The apparent ubiquity of the MMTV-beta DNA sequences in the genus Mus and the lack of variation in the pattern of restriction fragments containing these sequences within a species distinguishes them from MMTV-alpha. These results suggest that the MMTV-beta DNA sequences either are the evolutionary progenitors of the infectious MMTV genome or represent an accumulation of evolutionarily divergent MMTV-alpha insertions into the mouse germ line.

Animals↗
PubMed1982

Mapping the antigenic epitope for a monoclonal antibody against lysozyme.

A monoclonal antibody (HyHEL-5), prepared to chicken lysozyme c by the method of Köhler and Milstein, identified an antigenic site (epitope) that was shared by the lysozymes of seven different species of galliform birds. The lysozymes of two galliform species, bobwhite quail and chachalaca, shared only partial antigenic identity with the epitope defined by this antibody. Duck lysozyme did not react with the antibody at all. Amino acids that determined the epitope structure were tentatively identified by comparing the amino acid sequences of these lysozymes and assuming the antigenic changes produced by evolutionary substitutions are not due to long-range conformational changes. Arg 68 was identified as a determining amino acid. Arg 68 is hydrogen-bonded to Arg 45, and together these two amino acids form a basic cluster that may be a subsite of the epitope. The antibody inhibited lysis of Micrococcus lysodeikticus by chicken lysozyme. Additionally, Biebrich Scarlet, a dye that binds to the catalytic site, inhibited antibody binding to this lysozyme, which indicates that the epitope extends into the cleft region between Arg 45 and Arg 114. The epitope was hypothesized to involve a region measuring at least 13 x 6 x 15 A including the Arg 68-Arg 45 complex that borders the enzymatic catalytic site. Four other monoclonal antibodies to lysozyme have been partially characterized; each had a distinct pattern of binding specificity for various species of bird lysozymes.

Animals↗
PubMed1982

Genetics of the alpha 1,6-dextran response: expression of the QUPC52 idiotype in different inbred and congenic strains of mice.

Antibodies to dextran B512 were raised in various strains of mice and were assayed by a radioimmunoassay procedure. Idiotypic antibodies to the IgA(k) dextran B512 binding myeloma proteins QUPC52 and W3129 of BALB/c origin were prepared in rabbits. After adsorption each antiserum was specific for the immunizing myeloma protein and did not react with hundreds of other myeloma proteins; nonetheless, antibodies to dextran B512 from various strains of mice cross-reacted in these test systems. Of the 2 idiotypes tested, the W3129 idiotype was more universally expressed in different strains of mice. The QUPC52 idiotype was the predominant idiotype in BALB/c anti-dextran B512 antibodies and was found in only a few other inbred strains. Using a battery of congenic and inbred strains, it was shown that the QUPC52 idiotype was controlled by genes linked to the Igh complex locus (chromosome 12) and to the Ig kappa complex locus (chromosome 6). The W3129 idiotype was found in a number of stocks of mice in the genus Mus recently isolated from the wild. The QUPC52 idiotype thus far was found only in inbred mice.

Animals↗
PubMed1982

A cross-reactive idiotype, QUPC52 IdX, present on most but not all anti-alpha (1 replaced by 6) dextran-specific IgM and IgA hybridoma antibodies with combining sites of different sizes.

Seven BALB/c IgM, 4 BALB/c IgA, and 1 C57BL/6 IgA anti-alpha (1 replaced by 6) dextran hybridoma antibodies were characterized idiotypically. Five of the 7 IgM and all 4 BALB/c IgA proteins bear a cross-reactive idiotype present on the anti-alpha (1 replaced by 6) dextran BALB/c myeloma protein QUPC52 and on a majority of anti-alpha (1 replaced by 6) dextran antibodies in BALB/c mice. Of these 9 monoclonal antibodies, some have combining sites as large as 6 glucose residues, and some have combining sites as large as 7 glucose residues. Individual idiotypes present on QUPC52 are differentially expressed on the 9 hybridoma proteins that bear the cross-reactive idiotype. One BALB/c IgM hybridoma protein and the C57BL/6 IgA hybridoma protein did not react with anti-QUPC52 idiotypic antibodies; another BALB/c IgM hybridoma antibody showed only marginal reactivity.

Animals↗
PubMed1982

kappa-Chain restriction in anti-galactan antibodies.

Complete kappa-chain variable region sequences were determined for 10 light chains from anti-beta-(1,6)-D-galactan-binding monoclonal antibodies. Eight of these light chains were from IgM and two from IgG proteins. Seven of the eight IgM light chains were identical from position 1 to 95, the region encoded by the light chain variable region gene. The eighth differed at a single framework position. The two IgG light chains were identical in sequence and differed from the IgM light chains at two framework positions. All 10 light chains have an lle at position 96, as has been previously reported in 5 of 6 anti-galactan myeloma proteins. This residue is the first amino acid normally encoded by the light chain J gene. None of the germ line J genes encodes an lle at position 96, and furthermore the lle codon cannot be generated by alterations in the frame of recombination between the codon for amino acid 95 and J gene codons. These light chains are thus the product of an unusual and reproducible recombination event that appears to employ an extra codon at the 3' end of the variable region gene. Three different J segments are potentially used in these light chains J1 or J2 was found in three chains and J5 in seven.

Amino Acid Sequence↗