bluuebunny/crossref_metadata_2025 huggingface dataset:
Enhancing the frequency of antigen‐specific hybridomas
Authors: Fox Philip C. , Berenstein Elsa H. , Siraganian Reuben P.
Published: February 2007

Techniques were developed to enhance the frequency of antigen‐specific hybridomas by enriching for antibody precursor or ‐producing lymphocytes prior to cell fusion. Spleen cells of immunized animals were either (a) cultured in vitro for 3–4 days in the presence of the immunizing antigen or (b) adoptively transferred into X‐irradiated syngeneic recipients followed immediately by an in vivo antigen boost 4 days prior to use in cell fusion. There was a 10‐fold increase in the percentage of wells which develop antigen‐specific, antibody‐secreting hybridomas following these treatments. These techniques allow an increased yield of hybridomas with a greatly decreased technical effort.
Suppression of the primary IgM response by environmental teichoic acid
Authors: Oldfather John W. , Chorpenning Frank W.
Published: February 2007

It has been shown previously that ingestion of glycerol teichoic acid (GTA) in the conventional laboratory diet (8 mg/kg) is the stimulus for natural background responses to GTA in rats. Since injected GTA suppresses responses to sheep red blood cells (SRBC), it was suspected that dietary GTA also might be acting suppressively. A comparison of rats fed the conventional diet with rats fed a GTA‐free diet showed that ingested GTA markedly suppressed immune and background direct plaque‐forming cell (PFC) responses to SRBC. It appeared that a direct causal relationship existed between the degree of suppression and the amount of GTA exposure. When GTA‐deprived rats were force‐fed varying doses of GTA or when conventional animals were injected (i.p.) with GTA, increasing the total GTA dose resulted in decreased direct PFC responses to SRBC. Suppression was also observed when GTA‐deprived rats were force‐fed GTA‐containing Bacillus sp. ATCC 29726. The phenomenon of suppression by dietary GTA was not restricted to responses to SRBC, as similar results were obtained with chicken erythrocytes. When IgG PFC were measured, no difference between conventional and GTA‐deprived groups was observed. Thus, an IgM‐IgG shift does not seem to play a role in the mechanism of suppression by GTA.
Common neuroectodermal antigens on human melanoma, neuroblastoma, retinoblastoma, glioblastoma and fetal brain revealed by hybridoma antibodies raised against melanoma cells
Authors: Liao Shuen‐Kuei , Clarke Bryan J. , Kwong Pak C. , Brickenden Anne , Gallie Brenda L. , Dent Peter B.
Published: February 2007

The hybridoma system has been utilized to produce antibodies to characterize the cell surface antigens on human melanoma cells. On initial screening, two antibodies derived by the fusion of mouse myeloma cells (SP2/0‐Ag14) and splenocytes from a mouse immunized with a melanoma cell line (CaCL 78‐1) showed cross‐reactivity with 10 melanoma cell lines and did not react with any of 4 epithelial cancer lines, or 4 normal adult fibroblast lines. However, because of reactivity with 2 neuroblastoma cell lines, additional testing with other neuroectodermal derivatives was carried out and revealed a broad cross‐reactivity among melanomas, neuroblastomas, retinoblastomas and glioblastomas and against antigens shared by fetal but not adult brain. Thus, these results indicate the existence of common neuroectodermal antigens on melanoma cells. Before melanoma specificity can be claimed for, an antibody reactivity with nonmelanoma cells bearing these neuroectodermal antigen should be excluded.
Rabbits immunized against b6 allotype express similar anti‐b6 idiotopes
Authors: Roland Jaques , Cazenave Pierre‐André
Published: February 2007

Similar idiotopes are detected on anti‐allotypic antibodies synthesized by all rabbits immunized against the b6 allotype. The expression of similar idiotopes on anti‐b6 antibodies has been found in rabbits of all VHa allotypic types thus far studied. For their expression, these idiotopes require the interaction between VL and VH domains.
Antigen presentation and regulatory functions of human monocytes in the in vitro response of lymphocytes against purified protein derivative of tuberculin (PPD)
Authors: Treves Abraham J. , Tal Tova , Barak Vivian , Fuks Zvi
Published: February 2007

Monocyte functions in the primary and secondary proliferative responses of human lymphocytes against purified protein derivative of tuberculin (PPD) have been studied. By applying the antigen either directly to the responding cells or on antigentreated autologous monocytes, it was possible to distinguish between two different monocyte functions: antigen presentation and regulation. Their helper function in lymphocyte proliferation was found in cell‐free supernatants of autologous and allogeneic monocytes and was expressed better in the secondary rather than in the primary responses. The antigen‐presenting function of monocytes, however, was found to be more substantial in the primary response of lymphocytes. These results suggest that macrophage dependency of certain immune functions should be considered in regard to the various functions of macrophages, since at least quantitative differences for each of the macrophage functions were found in the primary and secondary proliferative responses.
Interferon enhances the amount of membrane‐bound β2‐microglobulin and its release from human Burkitt cells
Authors: Fellous Marc , Bono Rosa , Hyafil François , Gresser Ion
Published: February 2007

Human leukocyte interferon (HuIFN‐α) increased the amount of β2‐microglobulin on the surface of human Burkitt lymphoma cells (Ramos) and also increased the amount released into the culture medium. The effect was observed 1 h after addition of IFN. These results suggest that the increase in β2‐microglobulin on the cell surface of IFN‐treated cells is not due to a decreased shedding of antigen from the cell surface, nor an “unmasking” of surface antigen, but rather to an increased synthesis of antigen.
Experimental allergic encephalomyelitis in mice: encephalitogenicity of mouse central myelin basic proteins
Authors: Rauch Helene C. , Katar Malkhan , Montgomery Ilene N.
Published: February 2007

Two co‐extractable myelin basic proteins (MBP) were isolated and purified from mouse brain and designated: large (L), estimated to be composed of 160–164 amino acid residues, and small (S), estimated to contain 114–115 residues. The two proteins migrate separately in polyacrylamide gel electrophoresis (PAGE) in a pattern similar to rat MBP‐S and MBP‐L; mouse MBP‐L resembles rat MBP‐L, human, bovine and guinea pig MBP by PAGE and by amino acid analysis. This report demonstrates for the first time that mouse MPB‐L alone, and not mouse MBP‐S, is encephalitogenic for guinea pigs and mice.
The origin of IgA in chicken bile: its rapid active transport from blood
Authors: Rose M. Elaine , Orlans Eva , Payne Andrew W. R. , Hesketh Patricia
Published: February 2007

In the chicken, there are two bile ducts, one draining the left lobe of the liver directly into the duodenum and the other draining the right lobe via the gall bladder. Cannulation of these ducts enabled us to collect bile in unanesthetized birds and to compare the IgA concentrations of the hepatic bile, gall bladder bile and blood serum. Bile from the cystic and hepatic ducts of the same bird contained similar amounts of IgA (1.7 mg/ml), roughly 10 times as much as in serum, but considerably less than that found in the concentrated bile collected by aspiration from the gall bladder (8 mg/ml). Ligation of the two bile ducts resulted in a three to fourfold increase in the concentration of IgA in serum suggesting that IgA is normally removed from serum by the biliary route. This was confirmed by a substantial recovery of i.v. injected, radiolabeled monoclonal dimeric human IgA in the bile corresponding to a rapid active clearance from the blood circulation; negligible amounts of monomeric human IgA, similarly injected, were recovered from the bile.
Differentiation of thymocytes during chicken ontogeny: occurrence of a specific DNA ligase in relationship to cell size and surface antigens
Authors: David Jean‐Claude , Fedecka‐Bruner Barbara , Mishal Zohair , Vinson Denise , Rosenfeld Claude
Published: February 2007

The activities of two forms (8.2 and 6.2 S) of DNA ligase have been studied in chicken thymocytes at different stages of development (16 and 18 days of incubation, hatching and one month after birth). Thymocytes have been sorted out on the basis of size and T antigenicity (immunofluorescence) with the activated cell sorter. Three kinds of cells can be distinguished: (a) large, antigenically negative cells of 16 days of incubation, expressing only the “heavy” (8.2 S) form of enzyme. The activity of this form decreases during development and is very low after birth; (b) large, antigenically positive cells without DNA ligase activity and (c) small, antigenically positive cells, expressing the “light” (6.2 S) enzyme. This form of enzyme appears only from 18 days of incubation onwards. These results allow us to consider the 6.2 S DNA ligase activity as an additional marker (with cell reduction and T antigenicity) for thymocyte maturation.
Rat IgD myeloma protein: cell‐free translation of the δ mRNA and biochemical analysis of intracellular and membrane δ chain
Authors: Sire Joséphine , Alcaraz Gisèle , Bourgois Alain , Jordan Bertrand
Published: February 2007

The analysis of IgD molecules produced by the tumor‐derived IR 731 cell line showed that these molecules are not secreted and are only present at the cell surface as covalent dimers of 48000 dalton δ chains. The original IR 731 tumor secreted complete IgD; thus, the disappearence of secretion in the cell line was correlated with the absence of light chain synthesis.

The mRNA coding for the cell line δ chain was isolated and translated. The analysis of the translated δ chain (mol. wt.: 40000) confirmed the absence of a region having the size of a domain as previously shown by partial amino acid sequence. The observation that the absence in δ chain of this region, which comprises most of the CH2 domain, could be a general feature of murine species is discussed.
B cell heterogeneity. II. Transplantation resistance in xid mice which affects the ontogeny of B cell subpopulations
Authors: Quan Zoe S. , Dick Robert F. , Regueiro Benito , Quintáns José
Published: February 2007

F1 male offspring of CBA/N female mice carry an X‐linked immune defect, one manifestation of which is their inability to mount plaque‐forming cell (PFC) responses to certain thymus‐dependent (TD) and thymus‐independent (T1) antigens. Reconstitution of PFC responses in (CBA/N × BALB/c)F1 (NBF1) male mice with immature, immunocompetent neonatal liver cells requires preparative host irradiation. The need for irradiation is not absolute since the radiosensitive “barrier” can be overridden if a sufficiently large number of donor cells are transferred, or if one waits a sufficiently long period of time between transfer and challenge. An additional characteristic is that this resistance distinguishes between subsets of trinitrophenyl (TNP)‐specific and phosphorylcholine (PC)‐specific B cell progenitors.

Comparisons of lethally irradiated NBF1 females with males indicate that the two are qualitatively similar as hosts: responses to TD PC antigens appear before those to TI PC antigens, and quantal responses can be elicited under the appropriate conditions. In sublethally irradiated male recipients, however, the NBF1 male environment seems to operationally slow down the development of antigen‐reactive B cells; this is reflected as the transplantation barrier, which discriminates between compartments of B cell progenitors. It is precisely this differential delay of maturation which has enabled us to observe that the emergence of antigen‐specific responses proceeds in cycles of quantal, all‐or‐none, events. We discuss how the resistance of the immunodefective host environment to transplanted immature B cells may influence their differentiation.
Clonal regulation in the response to phosphorylcholine. II. Heterogeneity among T15 idiotype‐positive antibodies in inbred and wild mice
Authors: Pease Larry , Claflin J. Latham
Published: February 2007

Inbred stains of mice routinely respond to phosphorylcholine (PC)‐containing antigens, S. pneumoniae and PC‐keyhole limpet hemocyanin, with the production of three major families of antibodies. Two of these groups, the M 511 and M 603 idiotype (Id) families, are clearly heterogeneous, whereas the third, T15, normally exhibits much less heterogeneity and appears to be monoclonal in many strains. We report here on the occurrence of additional T15 Id+ antibodies which can be detected readily by isoelectric focusing. These antibodies have been observed in 9 of 12 strains examined at an average frequency of 10%. They occur in both the IgG1 and IgG3 isotypes. Collectively, the antibodies comprise a heterogeneous group although cofocusing antibodies were observed in individuals of a strain or between strains. Two H‐2‐congenic strains, BALB.B and BALB.G, regularly displayed multiple cofocusing T15 Id+ antibodies in each individual. Idiotypic analysis of these antibodies in all strains showed that each contained H, L and H‐L‐specific variable region determinants common to T15. A surprising finding was the consistent expression of a heterogeneous T15 Id response in two wild Mus musculus lines. Most of these antibodies cofocused with T15 Id+ antibodies in BALB.G or C3H as well as contained strain‐specific idiotypic determinants found among inbred mice. These data demonstrate that BALB/c and other inbred strains possess the genetic potential to generate a family of closely related T15 Id+ antibodies. The results further suggest that the restricted response usually seen among inbred strains is not normal and exists because of a highly selective regulatory system.
Idiotype‐anti‐idiotype interactions and the control of the anti‐β(2→6) polyfructosan response in the mouse: specificity and idiotypy of anti‐ABPC48 anti‐idiotypic monoclonal antibodies
Authors: Legrain Pierre , Voegtle Danielle , Buttin Gérard , Cazenave Pierre André
Published: February 2007

Seventeen hybridomas, secreting monoclonal anti‐idiotypic antibodies (IDA) directed against the BALB/c ABPC48 idiotype, were isolated from one immunized BALB/c mouse. Several IDA also bind another BALB/c idiotype: UPC10. ABPC48 and UPC10 are both myeloma proteins with a β(2→6)‐polyfructosan (levan) specificity. The binding of every IDA to the ABPC48 idiotype can be completely inhibited by levan molecules, but at different concentrations. Mutual inhibition assays between the IDA made it possible to define six groups of IDA which bind at least three different idiotopes of ABPC48.

Sixteen IDA have been studied by means of mouse anti‐anti‐idiotypic antibodies (Ab3) directed against two of them, IDA3 and IDA10. Anti‐IDA3 Ab3 recognize idiotopes particular to IDA3 which are not found on other monoclonal anti‐idiotypic antibodies (Ab2). Anti‐IDA10 Ab3 cross‐reacts with several monoclonal Ab2, including Ab2 with different spectrotypes belonging or not to the same isotype and Ab2 with different specificities for the ABPC48 idiotype.

Some IDA10 idiotopes are present in the polyclonal anti‐ABPC48 antibody response of BALB/c, A/J and CBA mice showing that they are recurrent and that their expression is not linked to a particular Igh‐C haplotype. In contrast, IDA3 idiotopes are not detected in the same anti‐ABPC48 antisera.
Enumeration of alloreactive helper T lymphocytes which cooperate with cytolytic T lymphocytes
Authors: Lutz Charles T. , Glasebrook Andrew L. , Fitch Frank W.
Published: February 2007

The number of alloreactive helper T lymphocytes (HTL) which cooperate with cytolytic T lymphocytes have been quantified in a sensitive limit dilution system. Approximately 1 in 330 splenic T lymphocytes developed helper activity in response to alloantigen of a single haplotype. Most HTL appeared to cooperate nonspecifically with the cytolytic cells. A significant number of alloreactive HTL precursors recognized H‐2 K or D alloantigen, but roughly 6‐fold more HTL precursors responded to H‐2 I, and roughly 20‐fold more responded to non‐H‐2, presumably Mls, alloantigen. These results support the hypothesis that alloantigen encoded by the H‐2 I and non‐H‐2 Mls regions preferentially stimulate helper T lymphocytes while alloantigens encoded by the H‐2 K and D regions preferentially stimulate cytolytic T lymphocytes.
Differences in the cell binding affinity of a cross‐reactive monoclonal anti‐Ia alloantibody in mice of different H‐2 haplotypes
Authors: Birnbaum Daniel , Kourilsky FrançOis M.
Published: February 2007

The analysis of an Ia specificity was performed in several H‐2 haplotypes by evaluating the affinity of the binding to mouse spleen cells of a monoclonal anti‐Ia alloantibody. We used the monoclonal antibody 10‐2.16 (Oi et al., Curr. Top. Microbiol. Immunol. 1978. 81: 115.) which recognizes on I‐A gene products a public specificity common to several H‐2 haplotypes (f, ja, k, r, s and u). One parameter of the affinity (dissociation rate constant) was determined by the kinetics of the dissociation of cell‐bound 125I‐labeled Fab fragments in the presence of homologous antibodies. The dissociation rate constants thus obtained were different when the antibody reacted with cells from mice bearing different haplotypes and were similar for reactions with different mouse strains bearing the same haplotype. These data strongly suggest that the Ia public determinants involved are not strictly identical in different haplotypes, and support the concept that a serological cross‐reaction does not necessarily mean structural identity between public antigenic determinants.
Phagocytosis and degradation of DNA‐anti‐DNA complexes by human phagocytes I. Assay conditions, quantitative aspects and differences between human blood monocytes and neutrophils
Authors: Lamers Marinus C. , de Groot Elisabeth R. , Roos Dirk
Published: February 2007

The uptake in vitro was studied of 3H‐labeled DNA‐anti‐DNA complexes by neutrophils and monocytes from human blood. Complexes were prepared from 3H‐labeled circular double‐stranded (ds) DNA of bacteriophage PM2 and anti‐dsDNA‐containing sera from patients with systemic lupus erythematosus. After phagocytosis, cells and medium were separated. The cells were treated with DNase to remove adherent and noningested complexes before the cell‐associated radioactivity was counted. Thus, only complexes inside the cells were measured. The medium was analyzed for acid‐precipitable radioactivity. In this way, we found that neutrophils only phagocytose the complexes, whereas monocytes phagocytose the complexes and degrade the antigen. In contrast, both types of phagocyte degraded the antigen in tetanus‐antitetanus complexes. The degradation took place after phagocytosis, inside the cells. The difference in DNA degradation between neutrophils and monocytes correlated with the difference in acid DNase activity of the lysosomal fractions: monocytes contained DNase activity, neutrophils did not. With complexes made from DNA with 131I‐labeled anti‐DNA, we found that both cell types degraded the antibody. Uptake of complexes and degradation of antigen increased with incubation time and cell concentration and was saturable with respect to complex concentration. The processes were inhibited by 5 mM mono‐iodoacetic acid or by low temperatures.
Major histocompatibility complex control of the idiotypic network in the MOPC 173 system
Authors: Boyer Claude , Schiff Claudine , Milili Michèle , Fougereau Michel
Published: February 2007

Regulation of the anti‐idiotypic response against MOPC 173 (M‐173, γ2a, ϰ myeloma of the BALB/c strain) was studied after syngeneic and allogeneic immunizations. The immunogen was M‐173 polymerized with glutaraldehyde, and hemagglutinating titers (HT) were determined with sheep red blood cell‐M‐173. H‐2 regulation of the response was clearly shown in mice having the same Igh‐Ca genes as the immunogen protein. H‐2d mice (BALB/c) are dispersed responders (HT: 80‐81 920) and 30% did not respond. H‐2k mice (BALB.K) are high responders (HT: 10240‐81920), and H‐2b mice (BALB.B) are nonresponders (HT < 40). In animals of a non‐BALB background, similar observations were made: Igh‐Ca, H‐2k mice (C58) were responders, whereas Igh‐Ca, H‐2b mice (C57L) did not respond. However, more dispersed HT values for the C58 animals as compared with those of the BALB.K suggest that a second level of regulation by background genes might be superimposed. When alloimmunization was used in strains not expressing the Igh‐Ca allotype (b, e, j), animals responded whatever the H‐2 haplotype (b, k, d). In this case, allotypic determinants might play a carrier role. All responder mice consistently made antiidiotypic antibodies as determined by hemagglutination and/or radioimmunoassay. Antibodies against new glutaraldehyde‐induced determinants were also produced but appeared in low amounts in syngeneic immunizations. In allogenic immunizations, various levels of anti‐allotype antibodies were detected.

BALB.B (and BALB/c) mice are responders to U‐10 (γ2a,ϰ, BALB/c myeloma), polymerized with glutaraldehyde and can respond to the M‐173 idiotype when they are immunized with M‐173 coupled to a heterologous carrier (a human γ1, ϰ myeloma protein, Kle). The unresponsiveness of BALB.B mice to M‐173 is thus idiotype‐specific and is not due to a defect in the anti‐idiotypic antibody repertoire. Injection of M‐173 before M‐173‐Kle prevents the response to M‐173 but not to Kle, which favors the occurrence of suppressive T cells.

Responsiveness is dominantly transmitted in (BALB.B × BALB.K)F1 mice which are Igh‐Ca/a, H‐2b/k heterozygotes. Unresponsiveness found with the Igh‐Ca, H‐2b combination is also observed in (BALB.B × B10)F1 which are Igh‐Ca/b, H‐2b/b. Since F1 animals between BALB.B and B10.A (3R), (4R) or (5R) are all nonresponders, genes involved in the response to M‐173 are not in the K, I‐A, I‐J, or I‐E subregions. A participation of the I‐B subregion or an I‐A + I‐E/C complementation cannot be excluded.
Immunoglobulin isotype expression II. Frequency analysis in mitogen‐reactive B cells
Authors: Benner Robbert , Coutinho Antonio , Rijnbeek Anne‐Marie , van Oudenaren Adrianus , Hooijkaas Herbert
Published: February 2007

The frequency of lipopolysaccharide (LPS)‐reactive B cells developing into clones that secrete the various immunoglobulin (Ig) classes has been determined in vitro, in cells from BALB/c mice, under culture conditions which detect all growth‐inducible cells. Secretion of the different Ig classes was assessed in the protein A plaque assay for Ig‐secreting, plaque‐forming cells by using developing antisera specific for either IgM, IgG1, IgG2a, IgG2b, IgG3 or IgA. In all lymphoid organs tested (spleen, bone marrow, mesenteric lymph nodes and thoracic duct), a considerable proportion of all B cells (5–20%) was induced by LPS to yield a clone of IgM‐secreting cells. Frequency determinations of LPS‐reactive cells giving rise to descendants secreting other Ig isotypes revealed that, on an average, and irrespective of the origin of the cells, 7% of all IgM‐secreting clones switched to the synthesis of IgG1, 39% to IgG2, 41% to IgG3 and 1% to IgA. Roughly the same frequencies of B cells switching CH gene expression were found among spleen cells of athymic nude mice. No correlation was found between the clonal frequencies of CH gene expression in polyclonally activated B cells and the in vivo “background” Ig‐secreting cells suggesting that the CH gene expression in B cells is influenced by the quality of stimulation and other regulating influences.
The tolerization of rat thymocytes to xenogeneic erythrocytes: kinetics of induction and recovery
Authors: Bunce Judith V. , Mason Donald W.
Published: February 2007

The intravenous injection of large doses of xenogeneic erythrocytes into rats completely abrogated the ability of thymocytes from such animals to provide help for antierythrocyte antibody responses in an adoptive transfer system. Thymocyte tolerance developed at a time when the thymocyte donors were producing antibody to the tolerizing antigen, and spleen helper T cells were not tolerized by the injection protocol. A minimum of two injections of 1 ml of packed erythrocytes, spaced 4–5 days apart, was required to induce loss of helper activity. Tolerance was fully developed within 16 h of the last injection of antigen. Helper activity recovered over a 24‐h period starting 5 days after this injection but could be delayed by giving further injections of antigen. These results suggest that the intrathymic pool of cells with assayable helper activity turns over in about one day and that precursors of this pool are probably also tolerizable. Suppressor cells could not be detected in tolerant thymocyte populations indicating that the loss of helper activity was either a consequence of clonal inactivation or that an antigen‐specific mechanism existed for the recruitment of helper cells from the thymus. The significance of these findings to the acquisition of self‐tolerance is discussed.
Characterization of the NPa idiotype through the analysis of monoclonal BALB/c anti‐(4‐hydroxy‐3‐nitrophenyl)acetyl (NP) antibodies
Authors: White‐Scharf Mary E. , Imanishi‐Kari Thereza
Published: February 2007

Spleen cells from BALB/c mice were fused with the nonsecreting myeloma line X63.Ag8.6.5.3 seven days after immunization with NP‐CG (4‐hydroxy‐3‐nitrophenyl) acetyl‐chicken γ‐globulin. The hybrid cell lines obtained were analyzed for heavy and light chain distribution, fine specificity, and idiotype.

The majority of monoclonal antibodies possessed either γ1 or μ chains. The distribution of L chains among these antibodies was approximately half λ and half ϰ. Thirteen monoclonal antibodies were grown as ascites tumors in mice. Examination of their fine specificity patterns showed that all of the λ antibodies are heteroclitic and have similar fine specificity patterns. Five of the seven ϰ antibodies are also heteroclitic, but their fine specificity patterns are more heterogeneous than those of the λ antibodies.

Polyspecific anti‐idiotypic sera directed against pooled primary serum antibody (R ā‐NPa) or against individual monoclonals were used for idiotypic characterization of the monoclonal antibodies. The R ā‐NPa bound all of the λ antibodies but none of the ϰ antibodies suggesting that the ϰ antibodies may be much more heterogeneous and were therefor not recognized in the presence of the more homogeneous λ antibodies. Further idiotypic analysis demonstrated that the λ antibodies, although no two are identical, are a very homogeneous group of antibodies which cross‐react with one another but not with the ϰ antibodies. Some, but not all, of the ϰ antibodies cross‐react with each other although none are cross‐reactive with the λ antibodies. Because the λ‐associated idiotype is recognized by the R ā‐NPa and its characteristics are similar to that of the C57BL/6 major idiotype (NPb), it is referred to as NPa. There may be a second major idiotype associated with at least some of the ϰ antibodies.
Analysis of an antigen‐specific, H‐2‐restricted cell‐free product(s) made by “I‐J−” Ly‐2 cells (Ly‐2 TsF) that suppresses Ly‐2 cell‐depleted spleen cell activity
Authors: Yamauchi Katsumi , Murphy Donal , Cantor Harvey , Gershon Richard K.
Published: February 2007

Immune Ly‐1−,2+ splenic T cells release material(s) (Ly‐2 TsF) into culture supernatants which (a) are antigen‐specific and (b) do not contain gene products of the I‐region of the MHC (major histocompatibility complex) (within the limits of our detection system) and which can lead to antigen‐specific suppression of spleen cell cultures depleted of all Ly‐2+ (Ly‐2 and Ly‐1,2) cells. The cells which make Ly‐2 TsF possess no detectable I region‐ or Qa‐1‐encoded determinants. Nonetheless, the ability of the material(s) to suppress target cells is restricted by genes closely linked to the MHC. No demonstrable evidence for a role of the polymorphic portion of genes linked to the Ig locus for this suppressive interaction to occur could be found. It is suggested that this material(s) may be the final effector moiety of the suppressor circuit because (a) it does not require the presence of any Ly‐2+ cells to work, and (b) in contrast to material(s) which induce suppression, there is no latent period; i.e. the Ly‐2 TsF suppresses both the early and late phases of the immune response in primary Mishell‐Dutton cultures. The MHC restriction that is imparted to the biologically active material is likely to be due to its “anti‐MHC” activity.

Since this is the first report in which the presence of I‐J‐controlled gene products was investigated on an antigen‐specific, Ly‐2 cell‐produced suppressor factor that worked in the absence of Ly‐2+ cells in the assay culture, the possibility that many, if not all, antigen‐specific suppressor effector cells and molecules are “I‐J−” must be considered.
Resistance of MRL/Mp‐lpr/lpr mice to tolerance induction
Authors: Amagai Takashi , Cinader Bernhard
Published: February 2007

Mice of strain MRL/Mp‐lpr/lpr develop resistance against tolerance induction with aggregate‐freed rabbit gamma‐globulin as early as 6 weeks of age and show very complete resistance when they are 10 weeks old; congenic mice MRL‐Mp‐+/+ of 6 and 10 weeks can be rendered tolerant. By reconstitution experiments, it was shown that T cells from MRL/Mp‐lpr/lpr are responsible for the resistance to tolerance induction. A cell which is not colchicine‐sensitive contributes to tolerance of MRL/Mp‐+/+, but not to MRL/Mp‐lpr/lpr mice. Resistance of T cells against tolerance depends on expression of the lpr gene.
Vaccination against autoimmune encephalomyelitis (EAE): Attenuated autoimmune T lymphocytes confer resistance to induction of active EAE but not to EAE mediated by the intact T lymphocyte line
Authors: Ben‐Nun Avraham , Cohen Irun R.
Published: February 2007

Autoimmune encephalomyelitis (EAE) can be induced in genetically susceptible rats by active immunization against myelin basic protein (BP) or by passive transfer of anti‐BP lymphocytes. We have developed in vitro lines of T lymphocytes reactive only against BP that produce EAE upon i.v. inoculation of syngeneic rats. The object of the present study was to learn whether attenuated anti‐BP line cells could be used to vaccinate rats against passive as well as active EAE. We inoculated rats i.v. with anti‐BP line cells that were attenuated by irradiation or treatment by mitomycin C. A single inoculation was sufficient to protect about 70% of rats from subsequent EAE induced actively. However, even repeated vaccination could not protect against EAE mediated by passive transfer of anti‐BP line cells. Thus, attenuated cells of EAE effector lines cannot vaccinate against the autoreactive effects of preformed effector T lymphocytes.
Transfer of memory cells into antigen‐pretreated hosts. II. Influence of localized antigen on the migration of specific memory B cells
Authors: Baine Yaela , Ponzio Nicholas M. , Thorbecke G. Jeanette
Published: February 2007

Following i.v. injection, 2,4,6‐trinitrophenol (TNP)‐primed memory cells localized in recipient lymph nodes draining a footpad injection of TNP‐hemocyanin (TNP‐KLH) in greater numbers than in contralateral nodes draining a p‐azobenzenearsonate‐coupled KLH injection. Such hapten‐specific, unilateral memory B cell localization was still observed in immunosuppressed mice when antigen injections were given as long as 4 days prior to the memory cell transfer. The memory cells could be challenged to form plaque‐forming cells by footpad injections of TNP‐labeled Brucella abortus at 5 days, but not one day, after cell transfer. The present studies further clarify some parameters of this adoptive memory, as a model for the study of persistent local memory. Measures that promoted the unilateral lymph node retention of 125I‐labeled antigen also facilitated unilateral accumulation of TNP‐specific memory cells. Such measures included pretreatment of the recipients with cyclophosphamide, rather than γ irradiation, injection of anti‐carrier antibody the day before antigen, or use of small doses of preformed immune complexes instead of antigen alone. In general, a high ratio of lymph node‐to‐spleen and lymph node‐to‐blood cencentration of antigen in recipients appeared crucial for unilateral localization of memory B cells. Splenectomy of recipients prior to cell transfer enhanced the difference in plaqueforming cell responses between draining and contralateral nodes, but decreased their difference when performed 1 day after cell transfer, suggesting that the spleen may have served as a trap for memory cells. I.v. injection of antigen at the time of B cell transfer also interfered with unilateral localization. The results demonstrate that in the presence of persisting depots of antigen within lympn nodes (and absence of significant amounts of antigen elsewhere), memory B cells can be retained locally without activation into antibody‐secreting cells. This mechanism may, therefore, be responsible for the phenomenon of local, humoral, immunological memory.
Genetic control of the expression of allelic Ig genes at the VHa locus in a1/a2 heterozygous rabbits
Authors: Gilman‐Sachs Alice , Dray Sheldon , Knight Katherine L.
Published: February 2007

Heterozygous rabbits representing 9 of 15 possible a1 and a2 heavy chain haplotype gene combinations among rabbits in the University of Illinois colony were analyzed for ratios of al to a2 in serum immunoglobulin (Ig). The Ig from rabbits of the a1x−y−n81f73g74de12,15 heavy chain haplotype in combination with any of three a2‐associated heavy chain haplotypes have higher ratios of a1 to a2 than Ig from rabbits in which a1 is encoded by 4 other heavy chain haplotypes. For example, the mean a1: a2 ratio for adult a1x−y−n81f73g74de12,15/a2x32y33,− n82,f71g75de12,15 rabbits was 12:1 compared to 5:1 for a1x−y33,30n83,f71g75de12,15/a2x32y33,− n82,f71g75de12,15 heterozygous rabbits. Family studies indicated that the a1:a2 ratio was under the control of the heavy chain chromosomal region or a locus closely linked to it. Whether the regulation is due to varying numbers of VH genes and/or J gene segments, a separate regulator gene, or more efficient joining of certain gene segments, has yet to be determined.
The immunosuppressive effect of monoclonal anti‐Lyt‐1.1 antibodies in vivo
Authors: Michaelides Mary , Hogarth P. Mark , McKenzie Ian F. C.
Published: February 2007

Monoclonal anti‐Lyt‐1.1 alloantibody was produced as tissue culture supernatant and administered to mice. The antibody, given intraperitoneally, resulted in the suppression of all T cell functions studied, but was without direct effect on B cells. Thus, skin and tumor allograft survival was prolonged and there was suppression of the delayed‐type hypersensitivity response; T cell help in the anti‐sheep red blood cell antibody response, responder cells in the mixed lymphocyte reaction (MLR), leucoagglutinin‐responsive cells, cytotoxic T cell (Tc) function and the induction of Tc were either totally or partially suppressed, all these responses being mediated by Lyt‐1+2− or Lyt‐1+2+ cells in CBA/H mice. By contrast, there was no inhibitory effect on the MLR‐stimulating or lipopolysaccharide‐responsive cells. The administration of the anti‐Lyt‐1.1 antibody was accompanied by a depletion of Lyt‐1.1+ T cells from both spleen and lymph node. These studies indicate that the monoclonal anti‐Lyt‐1.1 antibody is active in vivo with a selective effect on T cells. The results also have important implications for studies of T cell interactions in the mouse in vivo, and for similar studies in man.
Polymorphism of germ‐line immunoglobulin VH genes correlates with allotype and idiotype markers
Authors: Ben‐Neriah Yinon , Cohen Justus B. , Rechavi Gideon , Zakut Rina , Givol David
Published: February 2007

The polymorphic nature of the immunoglobulin VH genes was investigated by Southern blot analysis of liver DNA of sixteen different mouse strains and hybridization with VH probes. Differences in restriction enzyme pattern (REP) were observed and six different patterns of restriction fragments were found for the sixteen strains analyzed. No equivalent polymorphism was observed in another multigene family, the actins. The six patterns correlate with immunoglobin constant region allotypes (Igh‐1). Experiments with Igh‐1‐congenic strains suggest that the VH REP is linked to immunoglobulin constant region haplotype. Mouse strains which share inherited idiotypes also share identical VH restriction pattern. This provides a structural basis for the genetic linkage between idiotypes and allotypes. It also indicates that different strains carry different VH gene repertoires, which may be the basis for the expression of different inherited idiotypes in various strains. We propose that a VH group is a set of linked genes that are coinherited as a cluster with the constant region genes and that VH and CH can be regarded as an extended haplotype.
Functional activities of antibodies against brain‐associated T cell antigens II. Stimulation of T cell‐induced B cell proliferation
Authors: Jones Barry
Published: February 2007

B cells could be stimulated to proliferate by mitomycin C‐treated normal T cells that were activated by rabbit anti‐mouse brain antiserum (RaMBr). The B cell proliferation peaked 48 h after the initiation of the cultures. The T cell subset involved was Lyt‐1+, 2−. The B and T cells participating in the interaction did not need to be matched at the H‐2 locus. A long‐term T cell line (C.1.a), which was genetically restricted by the H‐2 complex in its antigen (ovalbumin)‐specific interaction with B cells, could also be induced by RaMBr to stimulate B cell proliferation. However, the RaMBr‐mediated interaction of C.1.a with B cells was genetically unrestricted. This suggests that T cell activation by RaMBr can bypass the requirement for the recognition of I region antigens, which is a feature of antigen‐specific interactions between T and B cells. In fact, RaMBr antibody appeared to function by bringing T cells into contact with B cells by the interaction of the B cell surface membrane Fc receptors with the Fc portion of RaMBr antibody bound to the T cell surface. Brain‐associated antigen(s) appeared to play a unique role in T‐B cell interactions because antibodies against other T cell surface antigens were inactive, e.g. those which remained in rabbit anti‐mouse thymocyte antiserum at a high titer after absorption with mouse brain tissue. Furthermore, the brain‐associated T cell antigen involved in the cellular interaction was differentiated from the Thy‐1 antigen by the failure of xeno‐anti‐Thy‐1.1 antisera to induce the interaction of B6.PL.Thy‐12 T cells with B cells, and the presence of activity in rat anti‐AKR mouse brain serum which did not contain antibodies against Thy‐1.1 or Thy‐1.2 determinants.

The data appear to support the hypothesis that the brain‐associated T cell antigen, as defined serologically in these studies, may represent a molecule which plays a role in the immunological functions of the Lyt‐1+, Lyt‐2− T cell subset.
Role of Ia.W39 in the interaction of antigen‐presenting cells with T and B lymphocytes
Authors: Huber Brigitte T. , Rosenwasser Lanny J.
Published: February 2007

Ia.W39 is a B cell differentiation antigen whose membrane expression is controlled by the xid gene. In this report we show that, analogous to its B cell expression, Ia.W39 is also present on a subset of Ia+ macrophages, indicating heterogeneity within that cell population.

The Ir gene(s) for the antigenic determinants on the A chain loop of beef insulin maps to the I‐Ab subregion of the H‐2 complex and, as we have previously reported, is associated with the private specificity Ia.W39. Depletion of Ia.W39+ macrophages eliminates their capacity to present beef insulin to immune T cells, whereas the presentation of the multideterminant antigen trinitrophenylated ovalbumin is reduced less than 50%. Furthermore, we found that H‐2b mice lacking Ia.W39+ cells are unable to make a secondary in vivo IgG plaque‐forming cell (PFC) response to beef insulin, while the primary IgG PFC response is not dependent on Ia.W39. No shift in the kinetics of the response, nor development of suppressor T cells could be detected in Ia.W39− mice, which would explain their apparent nonresponsiveness to beef insulin after boosting with this antigen. These results, therefore, may reflect a difference in the Ir gene control acting at the primary vs. secondary response level.
A subpopulation of small pre‐B cells in rabbit bone marrow expresses x light chains and exhibits allelic exclusion of b locus allotypes
Authors: Gathings William E. , Mawo Rose G. , Cooper Max D. , Young‐Cooper Glendowlyn O.
Published: February 2007

The expression of immunoglobulin heavy and light chains by pre‐B cells and B lymphocytes was examined by two‐color immunofluorescence in heterozygous b5b9 rabbits. Allelic exclusion of b5 and b9 x light chain allotypes was observed for both surface immunoglobulin‐negative pre‐B cells and surface immunoglobulin‐positive B lymphocytes. In newborn bone marrow, pre‐B cells and immature B lymphocytes expressing b9 were as numerous as those expressing b5. In contrast, circulating B cells and bone marrow plasma cells expressing the b5 marker outnumber b9+ cells by 2 to 1 in adult b5b9 animals. Whereas most B lymphocytes expressed x light chain b allotypes, approximately 80% of the μ heavy chain‐positive pre‐B cells did not. The pre‐B cells that expressed detectable light chains were relatively small lymphocytes. A model is presented which includes a “transitional” pre‐B cell that expresses both p chains and x chains.
Release of platelet‐activating factor (PAF‐acether) and leukotrienes C and D from inflammatory macrophages
Authors: Roubin Régine , Mencia‐Huerta Jean‐Michel , Benveniste Jacques
Published: February 2007

Macrophages (Mϕ) isolated from the peritoneal cavity of C57BL/6 mice were either untreated or treated with various eliciting agents (thioglycollate, sodium caseinate) or with an activating agent bacillus Calmette Guérin (BCG). The various populations were assessed for their ability to release platelet‐activating factor (PAF‐acether), an ether phospholipid mediator, and slow‐reacting substance (SRS), a lipoxygenase arachidonic acid derivative. PAF‐acether was recovered in higher amounts from BCG Mϕ, than from resident Mϕ, whereas elicited Mϕ, exhibited a marked decreased ability to release this mediator. Such variations were only quantitative as evidenced by the similar enzyme sensitivity and high pressure liquid chromatography (HPLC) retention times of the various PAF‐acether‐containing supernatants. Resident, BCG‐and sodium caseinate‐induced Mϕ released similar amounts of SRS, whereas thioglycollate Mϕ, exhibited once again a marked decreased ability to release this mediator. Comparing retention times on HPLC of resident and BCG Mϕ, SRS with those of synthetic leukotrienes C and D, molecular variations were noted. Even though both Mϕ, populations released higher amounts of leukotrienes C than D, the DIC ratio was higher in BCG Mϕ, than in resident Mϕ,. These results show that different environmental factors can influence the release of PAF‐acether and leukotrienes from Mϕ.
Suppression of T cell responses through competition for T cell growth factor (interleukin 2)
Authors: Günther Joachim , Haas Werner , Von Boehmer Harald
Published: February 2007

Addition of activated T cells or of cells from a cytotoxic T cell clone to mixed leukocyte cultures abolished the generation of cytolytic activity in these cultures. The cytolytic T cell response is completely restored if T cell growth factor (interleukin 2) is added, indicating that the suppression is due to absorption or consumption of IL2 by the added cells.
Production of immune interferon is regulated by more than one T cell subset: Lyt‐1,2,3 and Qat‐5 phenotypes of murine T lymphocytes involved in IFN‐γ production in primary and secondary mixed lymphocyte reaction
Authors: Landolfo Santo , Kirchner Holger , Simon Markus M.
Published: February 2007

Lymphocytes responsible for the production of IFN‐γ (immune interferon) in primary and secondary mixed lymphocyte reactions have been characterized with antisera specific for the Lyt‐1,2,3 and Qat‐5 alloantigens. A comparison was made between selected T cell subsets with respect to their ability to proliferate, generate cytolytic activity and produce IFN‐γ in response to H‐2 alloantigens.

The data indicate that (a) in primary mixed lymphocyte reactions, IFN‐γ is produced by Lyt‐1+, Qat‐5+ and by Lyt‐123+, Qat‐5+ T cells, (b) in secondary mixed lymphocyte reactions, an additional T cell subset, which is Lyt‐23+, Qat‐5−, participates in the generation of IFN‐γ and (c) the production of IFN‐γ does not correlate with either proliferation or the generation of cytotoxic lymphocytes.
Idiotypic relationship of anti‐Ia.2 antibodies
Authors: Grutzmann Rudi , Hämmerling Günter J.
Published: February 2007

Alloantiserum and hybridoma‐derived anti‐Ia antibodies were investigated for their idiotypic similarity using 4 monoclonal anti‐idiotypes raised against monoclonal anti‐Ia. With one anti‐idiotype, idiotypic cross‐reactivity could be found between three independently derived monoclonal BALB/c anti‐Ia antibodies, all of which were directed against the same serological determinant, Ia.2. The respective idiotype, tentatively designated la.2 idiotope, could also be demonstrated on anti‐Ia.2 antibodies in BALB/c alloantisera, but not on anti‐Ia against other specificities. The expression of the Ia.2 idiotope is linked to the Igh allotype; the use of Ig‐recombinant mice allowed us to map the Igh‐Ia.2 locus to the left of the T1S marker in the Igha V region.
Recruitment of functional subpopulations of T lymphocytes Initiator T lymphocytes recruit helper T cells for cytotoxic response
Authors: Livnat Shmuel , Corley Ronald B.
Published: February 2007

Initiator T (Ti) lymphocytes are defined by their ability to recruit other T cell populations in vivo In this study the function of T cells recruited into draining lymph nodes following injection of Ti cells primed to alloantigens in mixed lymphocyte culture was examined. The results demonstrate that alloantigen‐specific helper T cells that interact with cytotoxic T (Tc) lymphocyte precursors are recruited, as shown by the significantly higher frequencies of helper cells in draining lymph nodes compared with controls. However, neither Tc cells nor their precursors are recruited. Recruitment by Ti lymphocytes is therefore selective for certain T cell subsets. Proposals to explain the mechanism, specificity, and selectivity of recruitment are discussed. We suggest that Ti cells have a central role in both the initiation of T cell‐dependent immune responses and in the maintenance of immunologic memory. Their function is the rapid mobilization of T cell subclasses to a regional lymphoid organ where the immune response subsequently develops.