bluuebunny/crossref_metadata_2025 huggingface dataset:
Analysis of the repertoire of anti‐NP antibodies in C57BL/6 mice by cell fusion. I. Characterization of antibody families in the primary and hyperimmune response
Authors: Reth M. , Hämmerling G. J. , Rajewsky K.
Published: February 2007

Spleen cells from C57BL/6 mice sensitized to the hapten (4‐hydroxy‐3‐nitrophenyl)acetyl (NP) were hybridized with myeloma cells, and a variety of hybrid cell lines was isolated which secreted homogeneous anti‐NP antibodies. The antibodies were purified by affinity chromatography and their chain composition, affinity and fine specificity were determined. All antibodies recovered from the primary immune response carried λ light and μ or γ1 heavy chains. Their variable portions were nonidentical but similar in terms of hapten‐binding specificity with a higher affinity for the cross‐reacting haptens (4‐hydroxy‐3,5‐dinitro‐phenyl)acetyl (NNP) and (4‐hydroxy‐5‐iodo‐3‐nitro‐phenyl)acetyl (NIP) than for the homologous hapten NP. This heteroclitic property as well as the presence of λ, μ and γ1 chains are characteristic for primary anti‐NP sera of C57BL/6 mice.

In contrast, four families of anti‐NP antibodies, each with a characteristic fine specificity pattern, were found among the hybrid cell antibodies derived from the hyperimmune anti‐NP response. The antibodies of one of these families were related to the antibodies recovered from the primary immune response in that they were heteroclitic and carried λ light chains. All members of the other groups expressed k chains and were nonheteroclitic.

The finding of well‐defined antibody families in this system and the isolation of their members enable us to approach the problem of V gene expression and diversification in a new way.
Natural killer cells in guinea pig spleen bear Fc receptors
Authors: Arnaud‐Battandier F. , Bundy Bonita M. , Nelson D. L.
Published: February 2007

The phenomenon of natural cytotoxicity or spontaneous cell‐mediated cytotoxicity (SCMC) was investigated in guinea pigs and compared with two other in vitro cytotoxicity reactions: mitogen‐induced cellular cytotoxicity (MICC) and antibody‐dependent cellular cytotoxicity (ADCC). The same xenogeneic target cells were employed in all three cytotoxicity assays.

Organ distribution studies revealed that SCMC effector cell activity was present in spleen and peripheral blood but not in thymocytes. Bone marrow cells possessed low levels of SCMC effector cell activity. The organ distribution of effector cell activities for MICC and ADCC paralleled that for SCMC. Studies of the cell surface characteristics of the SCMC effector cell revealed that spleen cells nonadherent to antigen‐antibody but not to antigen‐F(ab')2 antibody‐coated surfaces possessed markedly reduced SCMC effector cell activity. In addition, spleen cells depleted of Fc receptor‐bearing cells by EA rosetting also possessed diminished SCMC effector cell activity, while cell populations enriched in Fc receptorbearing cells by EA resetting possessed enhanced SCMC effector cell activity. These fractionation techniques had similar effects on MICC and ADCC effector cell activity. Depletion of adherent spleen cells including macrophages by nylon wool column passage resulted in a population of cells with enhanced SCMC, MICC, and ADCC effector cell activity.

Thus, in guinea pig spleen the effector cells mediating SCMC were shown to belong to a population of nonadherent Fc receptor‐bearing lymphocytes possessing several cytotoxic capabilities including the potential of mediating MICC and ADCC.
Quantification of antigen‐reactive cells among human T lymphocytes
Authors: Van Oers M. H. J. , Pinkster J. , Zeijlemaker W. P.
Published: February 2007

The number of antigen‐reactive cells among human peripheral blood T lymphocytes was estimated by a limiting dilution analysis. Antigen‐induced lymphocyte activation was measured by means of incorporation of tritiated thymidine [3H]dThd. We have studied the frequency of memory T cells for the bacterial antigens tuberculin PPD and tetanus toxoid in immune donors, as well as the frequency of alloantigen‐reactive T cells.

In 11 different donors, the observed frequencies of the antigen‐reactive T cell ranged between 1:300 and 1:16000 for PPD; for tetanus toxoid values, between 1:750 and 1:11500 were obtained in 5 different donors. The frequency of alloantigen‐reactive T cells was found to be higher: between 1:200 and 1:600 (n=10). For 3 donors, the estimated frequencies proved to be reproducible over a period of several months. Finally, a correlation could be demonstrated between the frequency of PPD‐reactive T cells and the [3H]dThd incorporation of 4 × 104 PPD‐stimulated lymphocytes.
Importance of HLA‐D antigens for the cooperation between human monocytes and T lymphocytes
Authors: Hansen G. Sønderstrup , Rubin B. , Sørensen S. F. , Svejgaar A.
Published: February 2007

The ability of autologous and allogeneic monocytes to restore the responsiveness of purified T lymphocytes to tuberculin (PPD) was studied in twelve different experiments involving eleven unrelated T lymphocyte donors and a large number of related and unrelated monocyte donors. T lymphocyte suspensions were prepared by passage of peripheral blood mononuclear cell suspensions through Ig‐anti‐Ig‐coated columns. Monocyte‐enriched suspensions were prepared from mononuclear suspensions by density gradient centrifugation on albumin solution and added to the T cell suspension in a final concentration of 3%. Autologous monocytes and monocytes from related donors sharing one HLA haplotype with the T cell donor partly restored the responsiveness to PPD, and so did monocytes from unrelated donors who shared HLA‐D alleles with the T cells. None of eight monocyte suspensions from eight unrelated donors sharing no HLA‐D antigens with the T cell donor were able to restore the responsiveness to PPD. The difference between HLA‐D‐sharing and nonsharing monocytes could not be explained by a simultaneous allogeneic reaction, as the addition of monocytes autologous with the T cells to mixtures of non‐HLA‐D‐sharing T cells and monocytes partly restored the response. Moreover, experiments with the addition of PPD to ordinary mixed leukocyte cultures revealed only a moderate depression on the PPD response. It is concluded that in man – as in mice and guinea pigs – some HLA‐D identity between monocytes and T lymphocytes is necessary for a cooperation to take place in the secondary immune response. This observation emphasizes the homology between HLA‐D antigens in man and Ia antigens in animals.
Immune function in aged mice IV. Loss of T cell and B cell function in thymus‐dependent antibody responses
Authors: Callard R. E. , Basten A.
Published: February 2007

The decline in antibody‐forming potential with age was studied in mice by analyzing the relative contributions of T and B cells in collaborative PFC responses to the T cell‐dependent antigens, dinitrophenylated (DNP) keyhole limpet hemocyanin, DNP‐human IgG, fowl IgG and sheep red blood cells. The experimental system involved the adoptive transfer of purified lymphocyte populations derived from the spleens of old and young mice into young irradiated recipients. An unequivocal reduction in B cell function was demonstrated in both primary and secondary antibody responses. In the former, young T cells failed to restore responsiveness to old B cells, while in the latter, similar results were obtained, even if old B cells were primed in the presence of young T cells. Despite the lower level of antibody production in old mice compared with young, no decrease in the number of antigen‐binding cells (detected by resetting and autoradiography) was observed. Taken together, these results suggest that a defect exists in B cells which is independent of T cell help, and that it is not due to cell depletion but rather to a qualitative abnormality in the cells themselves.

Old T cells did not collaborate with young B cells as well as young T cells did, indicating a loss of helper T cell activity. Spleen cells from old mice, when mixed with young spleen cells at a 1:1 ratio, caused a reduction in their antibody‐forming potential which was consistent with the presence of cells with suppressive activity. The suppressor cells were shown to be T cells since their effect was abrogated by treatment with anti‐Thy‐1.2 serum plus complement and enriched by passage through nylon wool columns.

These findings emphasize the need to use purified cell populations in analysis of responsiveness in old animals and highlight the multifactorial nature of the mechanisms involved in the ageing process.
The insulin receptor as a universal marker of activated lymphocytes
Authors: Helderman J. H. , Reynolds T. C. , Strom T. B.
Published: February 2007

Although resting, rat thymus‐derived spleen lymphocytes do not bear insulin receptors, these receptors emerge upon the membrane of alloimmune rat T cells subsequent to skin grafting. These studies examine conditions that permit the emergence of lymphocyte insulin receptors upon T or B‐enriched rat lymphocyte populations. Allogeneic stimulation was accomplished in vivo by skin grafting from (Lewis × Brown Norway)F1 (LBN) to Lewis male rats or by Graft‐vs.‐host (GVH) reaction established by intraperitoneal injections of 2 × 108 Lewis leukocytes each week for a total of four injections into LBN animals. In vitro allogeneic stimulation was provided by one‐way mixed lymphocyte cultures between the same strains. Lastly, both T and B‐enriched populations were interacted with mitogens concanavalin A (Con A), phytohemagglutinin (PHA‐P), and lipopolysaccharide (LPS). The T‐enriched populations were highly enriched for T lymphocytes (90%) and macrophage‐depleted while B‐enriched populations contained nearly 85% B cells. Insulin receptors emerged upon T‐enriched cell populations consequent to allogeneic skin grafting with saturability, specificity and high affinity (kd = 1.1 nM) as well as after GVH. Responder strain lymphocytes developed an insulin receptor during allogeneic mixed lymphocyte cultures within 72 h of initiation. Specific insulin‐binding sites also appeared upon T‐enriched populations after culture with PHA‐P and Con A but not LPS. Conversely, B cells developed an insulin receptor after interactions with LPS but not PHA‐P or Con A. Anti‐Ig and complement but not rat anti‐T cell antibody abrogated the ability of LPS to induce an insulin receptor on B‐enriched cells. Binding specificity was demonstrated by the inhibition of [125I]‐labeled insulin in the order porcine insulin ≊ desalanine insulin > proinsulin ≫ desoctapeptide insulin with growth hormone exhibiting no such inhibition. These data demonstrate that a true insulin receptor appears upon both T and B cells consequent to activation. The lymphocyte insulin receptor is a universal marker of cellular activation as it is not restricted to clone or species and may be applied to B and T lymphocytes of mouse, rat and man.
Expression of HLA‐A, but not of HLA‐B, in mouse‐human somatic cell hybrids carrying the region p21→pter of human chromosome 6
Authors: Zmijewski C. M. , Engel E. , Croce C. M.
Published: February 2007

Mouse‐human somatic cell hy brids containing human chromosome 17 carrying the region p21→pter of human chromosome 6 and no other human chromosomes, were found to express HLA‐A but not HLA‐B. Counterselection of the hybrid cells in medium containing 5‐bromodeoxyuridine resulted in the growth of hybrid cells that have concordantly lost the expression of HLA‐A and the human translocation chromosome.
Stimulation of immune reactivity by methoxysubstituted glycerol ethers incorporated into the feed
Authors: Boeryd B. , Nilsson T. , Lindholm L. , Lange S. , Hallgren B. , Ställberg G.
Published: February 2007

In mice, the plaque‐forming cell response to sheep red blood cells was stimulated by a mixture of methoxy‐substituted glycerol ethers isolated from Greenland shark liver oil and by synthetic 1‐0‐(2‐methoxyhexadecyl)‐glycerol, given in the diet. In preliminary experiments, this synthetic compound also increased the ability of parental spleen cells to induce graft‐vs.‐host reactions in F1 hybrid mice. Glycerol ethers occur in the bone marrow fat of mammals and in the membrane phospholipids. It is postulated that the methoxy‐substituted glycerol ethers supplied in the diet may stimulate the bone marrow and/or may be incorporated into membrane lipids, thereby changing the structure and function of the membranes.
Induction of B lymphocyte colony growth in vitro by thymus‐derived stimulating factor
Authors: Sredni B. , Gopas J. , Rozenszajn L. A.
Published: February 2007

Murine lymphoid cells which were stimulated in liquid culture containing thymus culture fluid (Thy‐CF) and seeded in a soft agar culture system, proliferated and developed into B cell colonies. Two types of colonies were formed: large colonies within the upper layer and small flat colonies on the surface of the upper layer.

Thy‐CF prepared from cells of normal hydrocortisone‐treated mice had a higher cloning potential than Thy‐CF prepared from untreated mice. At concentrations of Thy‐CF in culture medium greater than 35%, Thy‐CF prepared from normal mice had an inhibitory effect on colony formation.

Cells of nude mice were also able to form B cell colonies if thymocytes of normal mice were mixed with lymphoid cells in the culture medium. Thymocytes elaborate a B lymphocyte colony‐stimulating factor which, with the help of T cells, triggers a B cell population into colony formation and immunoglobulin production.
Anti‐H‐Y responses of H‐2b mutant mice
Authors: Simpson Elizabeth , Gordon R. D. , Chandler P. R. , Bailey D.
Published: February 2007

Two strains of H‐2b mutant mice, H‐2ba and H‐2bf, in which the mutational event took place at H‐2K, make anti‐H‐Y cytotoxic T cell responses which are H‐2‐restricted, Db‐associated and indistinguishable in target cell specificity from those of H‐2b mice. Thus, alteration of the H‐2K molecule affects neither the Ir gene controlling the response, nor the associative antigen. On the other hand, one H‐2Db mutant strain, H‐2bo, although it makes a good anti‐H‐Y cytotoxic response, shows target cell specificity restricted to its own Dbo antigen(s), and neither H‐2b, H‐2ba or H‐2bf anti‐H‐Y cytotoxic cells kill H‐2bo male target cells. Thus, the alteration of the H‐2Db molecule does not affect the Ir gene of H‐2b mice, but it does alter the H‐2Db‐associative antigen.
Tissue distribution and quantitation of la‐like antigens in the rat
Authors: Mason D. W. , Gallico G. G.
Published: February 2007

The tissue distribution of a major histocompatibility complex (MHC)‐linked alloantigen in the rat was studied using antiserum raised by alloimmunization in a MHC‐congenic pair of rat strains. HO (Ag‐B5) rats were immunized with HO.B2 (Ag‐B2) skin grafts, and the tissue distribution of antigens recognized by the resultant antiserum was determined by cellular radioimmunoassay using different HO.B2 tissues as targets or immunoabsorbent. Linkage to the rat MHC was formally established by backcross analysis. The results obtained indicated that the tissue distribution of antigens recognized by this antiserum was closely similar to that reported for the distribution of Ia antigens in the mouse. Thus, T cells and thymocytes bound, respectively, 2% and 10% as much antibody as peripheral B cells while no binding to erythrocytes was detectable.

Quantitative immunoabsorption studies revealed that thymocytes and kidney tissue had, respectively, 20% and 2% of the absorbing capacity of spleen cells. However, both tissues were capable of completely absorbing out the anti‐B cell activity showing that they both possessed all the Ia‐like specificities detectable by this antiserum on peripheral B cells. A variety of procedures aimed at reducing the passenger leukocyte content of kidneys had only a slight effect on the absorbing capacity.

The cellular radioimmunoassay results indicated that on B cells, Ia‐like molecules are a major membrane component, present in amounts comparable with surface immunoglobulin molecules.

Studies at the single cell level, using a fluorescence‐activated cell sorter, indicated that the expression of Ia‐like alloantigens by rat thymocytes was very heterogeneous and that large thymocytes were the most heavily labeled. No Ia‐like alloantigens were detected on alloantigen‐activated T cells.

The findings are compared with published data on the mouse.
A new surface marker on mouse natural killer cells: Receptors for Helix pomatia A hemagglutinin
Authors: Haller O. , Gidlund M. , Hellström Ulla , Hammarström S. , Wigzell H.
Published: February 2007

Natural killer (NK) cells in normal mouse spleen have so far been found to lack conventional cell surface markers. Recently, neuraminidase treatment has been shown to uncover receptors for the A hemagglutinin of the snail Helix pomatia (HP) on populations (preferentially T cells) of human blood lymphocytes. We have used HP coupled to Sepharose in columns to fractionate mouse spleen cells. Fractions consisted of passed cells (fraction I) and cells eluted with the competitive hapten N‐acetyl‐D‐galactosamine at either 0.1 mg/ml (fraction II) or 1.0 mg/ml (fraction III). NK activity and percentage of T and B cells in each fraction were determined. B cells were found enriched in fraction I. T lymphocytes, among them alloreactive cytotoxic T cells (CTL), were eluted in fraction III. However, NK activity of normal CBA spleens was highest in fraction II, fraction III was somewhat enriched, whereas fraction I was depleted of lytic activity. Thus, while NK cells possess HP receptors, their binding properties seem to differ from those of CTL. HP receptors represent the first simple and reliable marker detected on NK cells and might be useful for purification of this cell type.
Preparation and characterization of antibodies to the λ chain variable region (Vλ) of mouse immunoglobulins
Authors: Ben‐Neriah Y. , Lonai P. , Gavish M. , Givol D.
Published: February 2007

The variable portion (VL) of the mouse myeloma protein MOPC‐315 (α, λ2) was obtained from its Fv fragment and was used to immunize rabbits. Anti‐VL antibodies were found to be specific to the VL region of protein 315 and to precipitate VL, Fv or light chain but not Fab' or intact M315. Anti‐VL 315 precipitates also the light chain of MOPC‐104E (μ, λ1) but not the intact protein. Intact immunoglobulins (Ig) containing λ chains, although they do not precipitate with anti‐VL, inhibit the binding of anti‐VL to VL in the radioimmunoassay. Radioimmunoassay inhibition studies demonstrated that Vλ1 and Vλ2 cross‐react extensively and that anti‐VL 315 can be used as a general anti‐Vλ reagent to detect molecules containing λ chains in mouse serum. Analysis of sera from several mouse strains indicated that Vλ‐containing molecules are present in approximately 3% of the Ig population, whereas the SJL strain has no detectable Vλ‐bearing molecules. In some of the antisera, anti‐VL 315 slightly cross‐reacted with VK‐bearing molecules. Anti‐VL 315 antibodies are not anti‐idiotypes since they react with myeloma proteins M315 (anti‐2,4‐dinitrophenyl MOPC‐104E (anti‐dextran) and HOPC‐1 (specificity unknown) and with most serum Ig containing λ chains. In view of the similarity of mouse λ chain sequences, it is suggested that most anti‐VL 315 is “anti‐framework” antibody.
Maternal allotype dominance and allelic exclusion in the B lineage cells of the newborn rabbit
Authors: Loor F. , Kelus A. S.
Published: February 2007

Newborn rabbits from parents which differ at the b allotype locus (K chain), show a strong maternal allotype dominance, especially in the bone marrow. This concerns not only the membrane but also the cytoplasm, and it cannot be simply due to a passive uptake of maternal immunoglobulins. The spleen precedes the bone marrow for a high level of expression of the paternal allotype. In the thymus, cells which contain cytoplasmic immunoglobulin are found at low frequency, but their absolute number represent a substantial contribution to the B lineage cell pool. A small proportion of such thymus cells do not show allelic exclusion of K chain allotypes.
Specificities in natural cell‐mediated cytotoxicity against lymphoblastoid cell lines I. Selective inhibition by cross‐competition
Authors: Koide Y. , Takasugi M.
Published: February 2007

The apparent nonselective effects of natural cell‐mediated cytotoxicity tested directly on different lymphoblastoid cell targets were found to be quite specific in the cross‐competition assay. The specificity was detected through inhibition of cytotoxicity by competitor cells sharing common specificities with the target cells. Cross‐competition tests were performed employing eight lymphoblastoid lines including four T and four B cells. Selective inhibition observed between lymphoblastoid cell lines indicated that T and B cell lines were more antigenically similar within each group than between them. The target antigens that distinguish T cell from B cell lines have been tentatively called TA‐T and TA‐B.
Time and antigen dose‐dependent variations of IgM antibody affinity
Authors: Neri R. , Pini C. , Vicari G. , Doria G.
Published: February 2007

Affinity of IgM antibodies elicited in mice by a single injection of dinitrophenylated dextran was measured by equilibrium dialysis. Maturation of affinity was found to occur with time, the rate of increase being higher after larger antigen doses. Maturation was followed by a decrease of affinity with time, the fall being more pronounced after lower antigen doses. These findings are more in line with the theory of antigen‐induced diversity rather than with the maturation theory.
Selective participation of immunoglobulin V region and major histocompatibility complex products in antigen binding by T cells
Authors: Lonai P. , Ben‐Neriah Y. , Steinman L. , Givol D.
Published: February 2007

Antigen‐binding inhibition studies using microscopic autoradiography were performed on T or B cell‐enriched lymphocyte populations. Antibodies specific for the “framework” of immunoglobulin heavy or light chain variable domains (VH or VL), or anti‐H‐2 and anti‐Ia antisera were used. T cell subclasses were separated with anti‐Lyt antisera and complement. It was found that antigen‐binding T cells of different subclasses can be inhibited selectively with only one of the two anti‐V region antibodies. Antigen binding to Lyt‐1+ cells was inhibited by anti‐VH, while Lyt‐2+,3+ cells were inhibited by anti‐VL specifically. Anti‐Ia antisera inhibited unprimed Lyt‐1+ antigen‐binding cells, whereas anti‐H‐2K or anti‐H‐2D antisera inhibited unprimed Lyt‐2+,3+ antigen‐binding cells, and both classes of immune T antigen‐binding cells.
Guinea pig immunoglobulin light chain isotypes II. The preferential expression of lambda chain‐bearing anti‐phenyltrimethylammonium antibodies is associated with restricted variable region expression, but not with any alteration in the proportion of lambda chain constant region isotypes, relative to normal immunoglobulins
Authors: Brunhouse R. F. , Cebra J. J.
Published: February 2007

The inbred strain 13 guinea pig expresses at least three isotypes of immunoglobulin (Ig) lambda (λ) chain. The proportion of these isotypes in normal Ig can be estimated by isolating a constant (C) region cyanogen bromide peptide from λ chains and quantitatively analyzing its component tryptic peptides. Anti‐phenyltrimethylammonium antibody, when analyzed in this way, was found to contain the same proportion of each λ chain C region isotype as did normal Ig.

The specific antibody light (L) chain pools were found to contain predominantly λ chains, although these are the minority type in guinea pig normal Ig L chain pools. Moreover, amino‐terminal, 5‐pyrrolidone‐2‐carboxylic acid‐containing peptides isolated from antibody λ chains show no demonstrable sequence heterogeneity, while their homologs isolated from normal λ chains are markedly heterogeneous.

That a restriction in heterogeneity of variable region framework residue positions is not associated with any alteration in the proportion of C region isotypes in these antibody λ chains, implies that translocation of all guinea pig λ chain variable region genes to the isotypic C region genes occurs in a random way.
Specific binding of poly(I) · poly(C) to the membrane of murine B lymphocyte subsets
Authors: Diamantstein T. , Blitstein‐Willinger Eveline
Published: February 2007

Indirect immunofluorescence revealed that 13% of BALB/c and 33% of CBA spleen cells of B type carry specific binding sites at their surface for double‐stranded poly(I)·poly(C). Pretreatment of BALB/c spleen cells with anti‐mouse immunoglobulin serum increased the number of B cells capable of binding poly(I)·poly(C) indicating the existence of a second B lymphocyte subpopulation carrying masked poly(I)·poly(C)‐binding sites. Pretreatment of the cells with mitogenic doses of either lipopolysaccharide (LPS) or single‐stranded polynucleotides, e.g. poly(I) or double‐stranded poly(A)·poly(U), failed to affect binding of poly(I)·poly(C) to the cells. Poly(I)·poly(C) converts small poly(I)·poly(C)‐binding lymphocytes into lymphoblasts carrying poly(I)·poly(C)‐binding sites. Lymphoblasts derived from LPS‐stimulated cells do not carry poly(I)·poly(C)‐binding sites. Thymocytes or splenic T cells failed to bind poly(I)·poly(C). As measured by thymidine uptake, CBA mice containing a higher percentage of poly(I)·poly(C)‐binding cells, are high responder mice to poly(I)·poly(C), compared with low responder BALB/c mice.
Cell‐mediated immunity to myelin basic protein in Lewis rats made unresponsive to experimental allergic encephalomyelitis
Authors: McGraw T. P. , Swanborg R. H.
Published: February 2007

Lewis rats with experimental allergic encephalomyelitis (EAE) exhibited cellmediated immunity to myelin basic protein as determined both with in vivo and in vitro assays. Positive skin test reactions and production of migration inhibitory factor (MIF) were observed before onset and after recovery from EAE. Rats rendered unresponsive to EAE exhibited in vitro cell‐mediated immunity to basic protein, although in vivo manifestations were depressed. However, tolerant rats failed to respond to the encephalitogenic determinant; rats with EAE exhibited cell‐mediated immunity to this region of the molecule. The results indicate that EAE‐unresponsive rats possess lymphocytes capable of responding to basic protein, but that reactivity to the encephalitogenic peptide is suppressed.
H‐2‐linked murine cytotoxic T cell responses specific for sendai virus‐infected cells
Authors: Kurrle R. , Röllinghoff M. , Wagner H.
Published: February 2007

CBA (H‐2k) mouse‐derived lymphochoriomeningitis virus and herpes simplex virus‐specific cytotoxic T lymphocytes lyse virus‐infected target cells compatible on either the H‐2K or H‐2D region. In contrast, CBA, C3H and AKR (H‐2k) mouse‐derived sendai virus‐specific cytotoxic T lymphocytes (CTL) fail to lyse H‐2D‐compatible virus‐infected cells. A similar lack of H‐2D region‐associated lytic activity was found with C57BL/6 and C57BL/10 (H‐2b) mice as well as with the recombinants B 10.A (2R) [Kk‐Db]and B10.A (4R) [Kk‐Db]. On the other hand, BALB/c (H‐2d) mice and A/J (H‐2a) mice do generate H‐2Dd‐associated sendai virus‐specific CTL. These results are in contrast to those obtained with (CBA x BALB/c)F1 and B10.HTT [Ks‐Dd] mice, which failed to mount Dd region‐associated CTL responses. It is concluded that D region‐associated sendai virus‐specific CTL responsiveness varies with the H‐2 genotype of the responder cells.
Thymic regeneration after dexamethasone treatment as a model for subpopulation development
Authors: Boersma Wim , Betel Ido , van der Westen Gerard
Published: February 2007

Three mouse thymocyte subpopulations with differing sensitivity to dexamethasone were characterized by size and density. Distribution patterns were determined for thymocytes during normal development and during regeneration after treatment with dexamethasone. From these, the subpopulation proportions were calculated and cell population kinetics were evaluated. Medium‐sized low density cells (type III) were least sensitive to dexamethasone and showed a relatively slow regeneration rate. The small cells of intermediate density, “early cortical” cells (type II), exhibited the most rapid regeneration rate, while the growth rate of the small cells of high density (type I) was intermediate. Cells of intermediate and high density were relatively sensitive to dexamethasone. The ratio of the phytohemagglutinin to concanavalin A (Con A) response increased after dexamethasone treatment with the increase in the proportion of type III cells. Con A responses were attributed to type II and type III cells. A tentative multipathway model for intra‐thymic T cell development is presented. Large cells, not confined to a certain trajectory in the density gradient, may represent a pool of proliferating cells of the other three types.
Ontogeny of susceptibility of mouse splenic B cells to tolerance induction in vitro by TNP‐D‐GL
Authors: Goldings Eliot A. , Mosier Donald E.
Published: February 2007

The susceptibility of mouse spleen cells to hapten‐specific tolerance induction of a primary in vitro thymus‐independent antibody response was examined. Both the induction of tolerance by 2,4,6‐trinitrophenyl‐D‐glutamic acid‐D‐lysine (TNP96D‐GL) and of antibody formation (elicited by TNP‐Brucella abortus) in neonatal spleen cell cultures were unaffected by anti‐Thy‐1.2 plus complement treatment. Spleen cells from neonatal mice were only slightly more sensitive to TNP96D‐GL tolerance induction than were cells from adult mice. The difference in susceptibility to tolerance induction was not nearly as great as that predicted by “clonal abortion”‐type theories of B cell tolerogenesis.
Reactivity of monoclonal anti‐melanoma antibodies with melanoma cells freshly isolated from primary and metastatic melanoma
Authors: Steplewski Zenon , Herlyn Meenhard , Herlyn Dorothee , Koprowski Hilary , Clark Wallace H.
Published: February 2007

Human melanoma cells, freshly obtained from nine primary and metastatic melanoma cases, were tested for binding of monoclonal anti‐melanoma antibodies produced in vitro by hybridoma clones. Monoclonal anti‐melanoma antibodies bind to melanoma cells but do not react with nonmalignant cells obtained from the same patients or with cells obtained from giant hairy nevus. These results confirm the existence of tumor‐specific antigens. Binding of monoclonal antibodies to melanoma cells of several origins, primary or metastatic, from different patients suggests the existence of tumor antigens shared by human melanoma cells. The binding pattern of different antibodies to various cells also predicts the existence of more than one tumor‐specific antigenic determinant on melanoma cells.
Interferon production in the murine mixed lymphocyte culture. I. Interferon production caused by differences in the H‐2 K and H‐2 D region but not by differences in the I region or the M locus
Authors: Kirchner Holger , Zawatzky Rainer , Schirrmacher Volker
Published: February 2007

Little use has been made previously of the observation that interferon is generated in the murine mixed lymphocyte reaction (MLR). We have found high levels of interferon in the supernatants of MLR between spleen cells of mice of different H‐2 type after one to two days of culture. Interferon production was also seen in strain combinations that differed only in the K or D region of the H‐2 complex. There was no production of interferon in strain combinations differing in the M locus or in the I region of the H‐2 complex. The latter combinations are known not to induce cytotoxic effector cells. Thus, there seems to be a dichotomy in allogeneic determinants between those of the I region and of the M locus inducing primarily lymphoprolifertion, and those of H‐2K and H‐2D inducing cytotoxic activity and interferon production.
The functional dissection of human peripheral null cells with respect to antibody‐dependent cellular cytotoxicity and natural killing
Authors: Ozer Howard , Strelkauskas Anthony J. , Callery Richard T. , Schlossman Stuart F.
Published: February 2007

A null lymphocyte subpopulation was isolated from human peripheral blood using a combination of Ficoll density gradient centrifugation, chromatographic separation of immunoglobulin‐bearing cells on Sephadex G‐200‐anti‐human F(ab'), columns and E rosette sedimentation. Null cell recovery averaged 5% of total peripheral mononuclear cells with greater than 90% purity by surface marker analysis. The purified null subpopulation was markedly enriched for antibody‐dependent cellular cytotoxicity (ADCC) and natural killing (NK) activity, while the Ig+′ and E' populations were virtually inactive. In the NK assay, the number of recovered lytic units in the null subpopulation was greatly increased over the total lytic units in unf ractionated cell populations, while enrichment in ADCC lytic units was strictly dependent on the concentration of null cells in effector populations. The null cell mediators of both ADCC and NK activity were shown to lack the Ia‐like surface antigen, p 23, 30. The results suggest that both ADCC and NK are mediated by the same population of null cells comprising less than 5% of peripheral lymphocytes and which lack the Ia‐like antigen p 23, 30, as well as mature T and B cell surface markers. The marked enrichment of NK activity in purified null populations suggests that E+ T cells may regulate NK by this subpopulation in vivo.
Specific binding of T lymphocytes to macrophages. IV. Dependence on cations, temperature and cytochalasin B‐sensitive mechanisms
Authors: Lipscomb Mary F. , Ben Sasson Shlomo Z. ‐ , Tucker Thomas F. , Uhr Jonathan W.
Published: February 2007

Peritoneal exudate lymphocytes (PEL) from immune guinea pigs adhere to macrophages carrying the relevant antigen and are thereby stimulated to proliferate in culture. The resultant PEL represent a population highly enriched with regard to their capacity to specifically rebind to antigen‐pulsed macrophages. We have studied the mechanisms underlying specific binding of lymphocytes to macrophages by examining the effects of physical and chemical modifications of the two cell types. Specific binding was inhibited by fixation of cells, metabolic inhibitors, low temperatures, cytochalasin B and divalent cation depletion. After specific binding has taken place, cation depletion, but not cytochalasin B or low temperatures, disrupts binding. These observations indicate that specific binding occurs by a series of discrete events that can be operationally distinguished.
Mitogenic activity of Mycoplasma pulmonis II. Studies on the biochemical nature of the mitogenic factor
Authors: Naot Yehudith , Siman‐Tov Rama , Ginsburg Haim
Published: February 2007

The mitogenic activity of Mycoplasma pulmonis has been demonstrated to reside in the membrane of this microorganism. Studies aimed at the identification of the membraneous mitogenic factor have revealed that membrane proteins are essential components of this mitogenic manifestation. In addition, it has been shown that the microorganism's outer surface membrane proteins are responsible for mitogenic activity. It has been shown, however, that isolated membrane lipids are not mitogenic for rat lymphocytes and are not required for the membrane's mitogenic potential.
Strain and sex dependence of carrier‐determined immunologic tolerance to guanosine
Authors: Borel Yves , Stollar B. David
Published: February 2007

Immunity and tolerance to guanosine were studied in male and female BALB/c and SJL mice and their hybrid and backcross offspring. Two assays were used to measure the primary and secondary immune response to guanosine keyhole limpet hemocya‐ nin: a Farr assay for circulating antibody and a hemolytic plaque assay for antibody‐ forming cells. SJL mice were higher responders than BALB/c mice. Although both strains of mice were rendered tolerant by guanosine‐isologous IgG, BALB/c mice were more susceptible to tolerance than SJL mice. Males were more easily tolerized than females in both strains of mice. This was best seen in the secondary response of mice made tolerant before primary immunization. Both male and female F1 mice were high responders like the SJL; nevertheless, both sexes were easily tolerized suggesting that tolerance is dominant. Results in backcross animals indicated that the genetic control was more complex than that of a single gene, and that it was neither linked to the gene of the immunoglobulin heavy chain nor to the H‐2 haplotype. Again, females were more resistant than males to tolerance induction. The significance of the sex difference in susceptibility to tolerance to nucleic acid antigens may be related to the female predominance in systemic lupus erythematosus.
Induction and characterization of isogeneic anti‐idiotypic antibodies to BALB/c myeloma S117: lack of reactivity with major idiotypic determinants
Authors: Seppälä Ilkka J. T. , Eichmann Klaus
Published: February 2007

Isogeneic anti‐idiotypic antibodies were induced by immunization of BALB/c mice with the BALB/c‐derived myeloma protein S117 which binds N‐acetylglucosamine‐containing antigens including Group A streptococcal carbohydrate (A‐CHO). Most BALB/c mice produced anti‐S 117 idiotypic antibodies, as shown by various different immunization protocols. The antibodies of individual mice were of intermediate to high affinity (2.8 × 106 M−1 to 1.4 × 108M−1). In isoelectric focusing, most individual antibodies were shown to consist of a small number of clonotypes, but each mouse produced its own unique set of clones so that the potential clonal repertoire of strain BALB/c is rather large. Most importantly, all isogeneic anti‐idiotypic antibodies were directed against idiotypic determinants of S 117 that are not shared with induced anti‐A‐CHO antibodies, whereas it has been shown previously that allogeneic and xeno‐geneic anti‐S 117 idiotypic antibodies react with idiotypic determinants unique to S 117 as well as with those that are shared with anti‐A‐CHO antibodies. The data suggest that the expression of S 117 idiotypic determinants in strain BALB/c is not under antibody‐mediated, anti‐idiotypic feedback control.
Immune response deficiency of BSVS mice. II. Generalized deficiency to thymus‐dependent antigens
Authors: Briles David E. , Perlmutter Roger M. , Hansburg Daniel , Little J. Russell , Davie Joseph M.
Published: February 2007

BSVS mice gave abnormally low IgG responses to 5 thymus‐dependent antigens as well as a weak delayed‐type hypersensitivity (DTH) response to sheep red blood cells. In contrast to IgG, the IgM antibody responses of these mice were normal to three T‐independent antigens as well as to all five T‐dependent antigens. The low immune responsiveness of BSVS mice was also reflected in the low levels of IgG2a, IgG2b and IgG3 in their normal serum. The low T‐dependent immune responses may result from BSVS mice having been selectively bred for susceptibility to infection with St. Louis encephalitis virus and Salmonella. C57BL/6J mice, which are also highly susceptible to Salmonella, gave low immune responses similar to, but genetically distinct from, those of BSVS mice. The levels of Ig‐positive and theta‐positive cells were normal in BSVS and C57BL/6J mice.
Antibody‐dependent cellular cytotoxicity against tumor cells. II. The promonocyte identified as effector cell
Authors: Domzig Wolfgang , Lohmann‐Matthes Marie‐Luise
Published: February 2007

Macrophage precursor cells were cultured from the bone marrow of mice in a liquid culture system in the presence of conditioned medium. To separate their different maturation stages, they were passed through a discontinuous Ficoll density gradient, treated with iron plus magnet or passed through glass bead columns. The different maturation stages have been tested for their function in antibody‐dependent cellular cytotoxicity (ADCC) against tumor target cells and in lymphokine‐induced macrophage‐mediated cytotoxicity. It is shown that the promonocyte, a nonadherent, nonphagocytic precursor cell, is a highly potent cytotoxic effector cell against antibody‐coated tumor targets but is totally inactive as an effector cell in lymphokine‐induced macrophage‐mediated cytotoxicity. In contrast, in the lymphokine‐induced macrophage‐mediated cytotoxicity the cytotoxic effector cell is a mature macrophage. Thus, ADCC seems to be a function of the macrophage precursor promonocytes, whereas lymphokine‐induced cytotoxicity is performed by mature macrophages. The relationship of promonocytes and killer (K) cells in ADCC against tumor targets is discussed.
Genetic control of the immune response to the terpolymer L‐glutamic acid60‐L‐aIanine30‐L‐tyrosme10 (GAT): II. Characterization of a cross‐reactive idiotype associated with anti‐GAT antibodies from responder and nonresponder mice
Authors: Thèze Jacques , Sommé Gérard
Published: February 2007

An anti‐idiotypic antiserum specific for the antibodies directed against the terpolymer poly(Glu60, Ala30, Tyr10), referred to as GAT, has been prepared in rabbit immunized with anti‐GAT antibodies purified from ascites fluid of BALB/c GAT responder mice. The specificity of this serum has been investigated. The interaction between 125I‐labeled anti‐GAT antibodies and the anti‐idiotypic antiserum is specifically inhibited by anti‐GAT antiserum or anti‐GAT ascites fluid. GAT and the copolymer of L‐glutamic acid50‐L‐tyrosine50, but not the closely related copolymer of L‐glutamic acid60‐L‐alanine40, inhibit the idiotype‐anti‐idiotype binding indicating a close association of idiotypic determinants with the antibody combining site. This GAT idiotype was found in the anti‐GAT antibodies of all individuals of all inbred strains of mice tested regardless of their allotypic markers. This GAT idiotype has also been found in serum of nonresponder mice immunized with GAT bound to methylated bovine serum albumin. Therefore, the existence of a GAT cross‐reactive idiotype represented in a very large number of inbred strains of mice has been identified. These results are in contrast to results obtained in other experimental systems where linkage was always observed between allotypic and idiotypic specificities.
Mac‐1: a macrophage differentiation antigen identified by monoclonal antibody
Authors: Springer Timothy , Galfré Giovanni , Secher David S. , Milstein Cesar
Published: February 2007

We have previously described the derivation of M1/70, a hybrid myeloma line secreting monoclonal rat anti‐mouse cell surface antibody (Springer, T., Galfre, G., Secher, D. S. and Milstein, C, Eur. J. Immunol. 1978. 8: 539). We have now investigated the cellular distribution of this antigen using a 125I‐labeled anti‐rat IgG indirect binding assay, the fluorescence‐activated cell sorter, autoradiography and precipitation of cell surface molecules. Screening with a tumor cell panel showed strong reactivity with a macrophage‐like line but no reactivity with B or T lymphoma lines. In normal tissues, M1/70 antigen was found to be present in small amounts on spleen and exudate granulocytes and a subpopulation of bone marrow cells, in moderate amounts on spleen and blood monocytes and expressed in much larger amounts on spleen histiocytes and peritoneal exudate macrophages. In contrast, M1/70 antigen was found to be absent from erythroid and lymphoid cells. M1/70 antibody precipitated two polypeptides of 190 000 and 105 000 mol. wt. which were present in much greater amounts on peritoneal exudate macrophages than on spleen cells. The expression on phagocytes of two other antigens identified by monoclonal antibodies M1/69 and M1/9.3 was also examined. Monocytes and granulocytes expressed large amounts of M1/69 and low amounts of M1/70 antigen, while in peritoneal exudate macrophages this pattern was dramatically reversed. M1/70 thus defines a differentiation antigen on mononuclear phagocytes and granulocytes, the expression of which is specifically increased during monocyte maturation. This antibody is the first to be described which recognizes a discrete molecule specific to phagocytes.
Role of adherent T cells and of B cells in the immune response to purified protein derivative of tuberculin
Authors: Arala‐Chaves Mario P. , Porto Maria T. , Hope Lapsly , Fudenberg H. Hugh
Published: February 2007

The role of monocytes, B cells, and adherent and nonadherent T cells in the response of purified protein derivative of tuberculin (PPD) as measured by [3H]thymidine uptake in vitro has been explored. The response to PPD was found to be highly dependent on monocytes, to approximately the same extent as previously found for the responses to concanavalin A (Con A) and phytohemagglutinin. The response to PPD was also found to be highly dependent on a population of adherent T cells different from the adherent T cell population involved in the response to Con A. In the case of PPD, the effect was additive, as opposed to the potentiating effect seen for Con A. Further, the adherent T cells involved in the response to PPD were much more sensitive to hypotonic shock than those in the response to Con A. B cells were also found to be important in the response to PPD, although only to a slight extent.
The in vitro migration of murine fetal liver cells to thymic rudiments
Authors: Pyke Kenneth W. , Bach Jean‐Francois
Published: February 2007

The migration of murine fetal liver cells to thymus rudiments was studied in vitro using a migration under agar technique. There appeared to be a minor population that migrated specifically to the thymus from the age of 10 to 14 days of gestation. The specificity of migration was demonstrated in 12‐day fetal liver cells by a series of competition studies. The ability of these cells to colonize a thymus rudiment was shown by further incubation after invasion of the epithelial thymus rudiments: small colonies of lymphoid cells were present in invaded tissue but absent from uninvaded control tissue. At 13 to 14 days of gestation, there appeared an additional population that migrated specifically to the spleen, as demonstrated, again, with a competition protocol. Studies with avian and human tissue as attractants in the same system showed that migration was specific to the thymus and did cross species barriers. This observation was used to demonstrate a similar attractant activity in cell‐free conditioned medium from human thymus epithelial cultures, and to demonstrate the absence of such a cell‐attractant factor in the conditioned medium from the thymus of a child with previously documented severe combined immunodeficiency disease.
Isolated hapten‐binding receptors of sensitized lymphocytes. IV. Expression of immimoglobulin variable regions in (4‐hydroxy‐3‐nitrophenyl) acetyl (NP)‐specific receptors isolated from murine B and T lymphocytes
Authors: Cramer Matthias , Krawinkel Ulrich , Melchers Inga , Imanishi‐Kari Thereza , Rajewsky Klaus , Ben‐Neriah Yinon , Givol David
Published: February 2007

Primary anti‐NP [(4‐hydroxy‐3‐nitrophenyl)acetyl] antibodies and NP‐specific receptors isolated from T cells of mice carrying the Ig‐1b allotype express the NPb idio‐type and a fine specificity marker (heteroclicity) which are both controlled by V genes in the heavy chain linkage group. While in antibodies both markers are invariably expressed in association with Ig light chains of the Λ type, antigen‐binding T cell‐derived receptors appear not to follow this rule in that they do not significantly bind to anti‐Λ antisera.

Mice of the SJL strain also carry the Ig‐lb allotype but express a defect in Λ chain synthesis, and their primary anti‐NP response is thus almost completely devoid of Λ‐bearing heteroclitic molecules of the NPb idiotype. In contrast, both NPb idiotype and heteroclicity are expressed in the majority of T cell‐derived receptors. This result permits a positive discrimination between T cell‐derived receptors and humoral antibodies in the SJL strain and shows that V gene expression follows different rules in the T and the B cell compartment.

The expression of Ig V regions in isolated T cell‐derived receptors was also analyzed with antisera binding to presumed “framework” determinants of the VH and the VΛdomains of Ig. A reaction of anti‐VH antiserum with isolated T cell receptors could clearly be demonstrated. The anti‐VΛ antiserum may bind to a minor fraction of the NP‐specific molecules in our T cell receptor preparations.