bluuebunny/crossref_metadata_2025 huggingface dataset:
Antigen recognition. III. Effect of phytomitogens on antigen‐receptor capping and the immune response in vitro
Authors: Lee K.‐C. , Langman R. E. , Paetkau V. H. , Diener E.
Published: February 2007

Concanavalin A (Con A) has been shown to act differently on bone marrow‐derived (B) cell responses to antigen in vitro depending on the presence or absence of thymus‐derived (T) cells. When T cells were present, Con A suppressed the immune response to polymeric flagellin (POL) of Salmonella adelaide (a T‐independent antigen) and sheep erythrocytes (SRBC) (a T‐dependent antigen). In contrast, when T cells were absent, Con A enhanced the anti‐POL response and allowed an otherwise very low anti‐SRBC response to return to normal levels. Con A inhibited POL‐induced capping in a manner that was apparently independent of the presence of T cells. In a number of cases, the effects noted with Con A were also found with another mitogen, phytohemagglutinin. Some possible implications of these findings on the mechanism of T‐B cell collaboration and the relationship between capping and immune induction are discussed.
Localization of lymphocytes producing macrophage migration inhibitory factor in albumin density gradients
Authors: Pick E.
Published: February 2007

Lymph node cells of guinea pigs immunized with complete Freund's adjuvant were fractionated by centrifugation on a discontinuous albumin gradient and the resulting cell layers tested for their ability to release macrophage migration inhibitory factor (MIF) upon in vitro exposure to tuberculin purified protein derivative. It was found that MIF production is limited to lymphocytes of lower density, representing only one fourth of all cells, while high density, small lymphocytes are inactive in this respect. Peak MIF production was usually associated with top layer (lowest density) cells, cultured at a concentration 5–10 times lower than that required for MIF formation by unfractionated cells.
Hapten‐specific augmentation of the anti‐idiotype antibody response to hapten‐myeloma protein conjugates in mice
Authors: Janeway C. A. , Paul W. E.
Published: February 2007

BALB/c and CBA mice produce little or no anti‐idiotype antibody upon immunization with IgG (γ2a) myeloma proteins derived from BALB/c mice. Similarly, DNP (2,4‐dinitrophenyl) conjugates of such proteins are poorly immunogenic in these mice and give rise to little anti‐DNP or anti‐idiotype antibody. Preimmunization of such mice with a DNP‐protein, such as DNP‐OVA (ovalbumin), prepares these animals for substantial anti‐idiotype responses to DNP conjugates of the BALB/c γ2a myeloma protein LPC‐1. This may be defined as a hapten‐specific helper effect.

However, transfer of lymphoid cells from DNP‐OVA primed mice into irradiated syngeneic hosts does not prepare the recipients for an anti‐idiotype response and, thus, the effect does not appear to be mediated by DNP‐specific helper cells. By contrast, the transfer of serum from DNP‐OVA primed mice into normal recipients or of immune serum together with normal cells into irradiated hosts prepares them for anti‐idiotype responses to DNP‐LPC‐1. The active component of the immune serum appears to be an anti‐DNP antibody, as it can be removed by passing the serum over agarose columns to which either a DNP‐protein or an anti‐mouse‐κ antibody has been linked. The augmenting antibody appears not to function by inhibition of antigenic competition, as both anti‐DNP and anti‐idiotype antibody responses are increased by infusing such antibody.
The sensitivity of antibody to 2‐mercaptoethanol treatment in three Australian marsupials
Authors: Yadav M.
Published: February 2007

Immunological responses to injections of sheep erythrocytes, Salmonella adelaide flagella and bacteriophage ϕX174 were investigated in three species of Western Australian marsupials (Setonix brachyurus, Macropus eugenii and Trichosurus vulpecula).

Two classes of immunoglobulins similar to the eutherian IgM and IgG were normally formed to the antigens. In addition, a low molecular weight anti‐phage ϕX174 antibody of about 80 000 daltons was present in the three species. In Setonix the 19 S antibody converted to 7 S type relatively slowly in the primary response, but rapidly in the secondary response. The 2‐mercaptoethanol treatment showed that both 19 S and 7 S antibody was sensitive in variable proportions ranging from 0 to 100% and a different lability was noted to each of the antigens in the three species.

In general, results indicated that the immune response in the marsupials did not lack the important features which typify an immune response in eutherians.
Experimental autoimmunno‐oophoritis in the rat
Authors: Janković B. D. , Marković B. M. , Petrović Spomenka , Isaković Katarina
Published: February 2007

Lesions were produced in the ovaries of Wistar rats by means of a mixture of bovine ovary and complete Freund's adjuvant. The inflammatory infiltrates consisted of lymphocytes and histiocytes. Delayed skin hypersensitivity to ovarian tissue was followed by the appearance of anti‐ovary antibody in the circulation.
Genetic control of the tendency to tolerance circumvention
Authors: Rose J. E. M. St , Cinader B.
Published: February 2007

Random bred rabbits were injected with human serum albumin (HA) at birth to induce tolerance to HA. Tolerant animals were then immunized with a p‐azobenzenesulfonic acid derivative of human albumin (HA‐D31). A proportion of these animals, 42 %, (27/64) responded with the production of antibody to HA‐D31 (TCR‐responders). Matings were carried out between TCR‐responders and between TCR‐nonresponders, and the offspring were subjected to the same procedures as their parents. Selective breeding of TCR‐responders produced 70 % (12/17) responders after two generations. Selective breeding of TCR‐nonresponders produced only TCR‐nonresponders after the third generation. Fourth generation offspring of TCR‐nonresponders, which were not injected with HA at birth, produced an antibody response to HA‐D31. The specificity of this antibody was analyzed by agglutination‐inhibition tests and was found to be similar to that of antibody produced in unselected rabbits. The genetic control of responsiveness to HA‐D31 in HA‐tolerant rabbits is polygenic and autosomal.
Isolation of f(c)5μ and fabμ fragments of human igm
Authors: Zián J. , Bennett J. C.
Published: February 2007

It has been shown previously that preparation of F(c)5μ and Fabμ fragments from IgM is limited both by low yields and incomplete cleavage and that digestion with hot trypsin is superior to papain in the absence of cysteine. The present study demonstrates that by using trypsin digestion, F(c)5μ fragment is relatively much more stable than Fabμ fragment. Optimum conditions for splitting with trypsin were 60°C for 20 min.

Sedimentation constants of F(c)5μ and Fabμ fragments were 10.9 and 3.7 S, respectively. Molecular weights derived from polyacrylamide disc electrophoresis in sodium dodecyl sulfate were 340 000 and 48 000 for F(c)5μ and Fabμ respectively. The molecular weight of F(c)5μ fragment falls to 33 600 after partial oxidative sulfitolysis.

Amino acid and sugar composition of the fragments were determined and accounted for approximately 85 % of the IgM molecule. That portion which was lost is most likely a glycopeptide cleaved from the hinge area.
Immunosuppression with busulphan: the effect on spleen, marrow and thymus cells of mice
Authors: Addison I. E.
Published: February 2007

The immunosuppressive activity of busulphan, an anti‐tumor agent, was studied using cell‐transfer experiments. Spleen cells from busulphan‐treated donors were many times less active than normal cells, while normal cells could restore competence to busulphan‐treated mice. It was concluded that the drug acts directly upon immunocompetent cells, but has no effect upon the environment in which they work. Further experiments showed that spleen from busulphan‐treated donors did not show synergy with normal marrow or thymus cells, and that neither marrow nor thymus cells from these donors show synergy with the reciprocal normal cell. It was concluded that busulphan affects both thymus‐dependent (T) and thymus‐independent (B) cells. However, marrow alone restored competence to busulphan‐treated recipients which, therefore, must contain an excess of T cells. This may be because mice normally contain an excess of T cells, or because there is a subpopulation of T cells outside the thymus and spleen, unaffected by busulphan.
Genetic control of phytohemagglutinin responsiveness by murine spleen cells and correlation between the release of lymphotoxins and dna synthesis
Authors: Donner Mireille , Vaillier D. , Burg C.
Published: February 2007

Response to phytohemagglutinin was investigated in adult mouse spleen cells of congenic strains which differ only at the H‐2 locus. The stimulation of spleen cells was assessed by release of lymphotoxins and DNA synthesis.

The results obtained show that the response to PHA markedly varied according to the genetic origin of spleen cells. Cells with H‐2a and H‐2b specificities responded better to PHA than those with H‐2f and H‐2s specificities. Furthermore, there was good correlation between the amount of released lymphotoxins and the level of DNA synthetic response.

These results are discussed on the basis of an H‐2 linked genetic control of PHA response in mouse strains.
Antigenic strength investigated by cell‐mediated lympholysis in mice
Authors: Abbasi K. , Festenstein H.
Published: February 2007

The cell‐mediated lympholysis (CML) technique has been used to investigate antigenic strength in mice. Investigations of the parameters of the technique led to calculation of specific chromium release on the basis of the effect of 5‐day mixed lymphocyte cultures (MLC) incubated for 4 h with 3‐day 51Cr‐labeled phytohemagglutinin (PHA) transformed blast cells in a ratio of 50:1.

The specificity of the reaction was established in mirror experiments and both major histocompatibility locus and M‐locus (non H‐2) incompatible MLC were tested against the appropriate PHA blasts. While strong MLC stimulation occurred in both these types of incompatible combinations, a specific killing capacity for CML developed only in the former. Furthermore, M‐locus gene products per se do not seriously influence the outcome of allograft survival a result consistent with those of others working in the rat and man, in which species the capacity to respond in MLC and to kill in CML and/or to reject allografts can be separated.

The fact that M‐locus stimulation does not lead to cell killing could be due to the absence of a concomitant serologically defined incompatibility; if so, the serologically undetectable lymphocyte activating determinants of the major histocompatibility system linkage group could have a similar biological function to that of the M‐locus.
A familial incidence of structurally restricted antibodies to pneumococcal polysaccharides
Authors: Pincus J. H. , Mage Rose G. , Alexander C. , Chace Nina M.
Published: February 2007

Rabbits were selected for their ability to produce structurally restricted anti‐polysaccharide antibodies when immunized with either type III or type VIII pneumococcal vaccines. Close relatives and offspring of these animals also exhibited this trait, indicating that it is inheritable. Within two groups of such restricted responders the trait could be observed with either type of vaccine. No correlation was observed with the magnitude of the immune response and the degree of antibody restriction. Furthermore, there was no correlation of this trait with sex, nor was there any obligate association with any a or b locus allotype. Isoelectric spectra of antibodies produced by individual rabbits showed no gross similarities, although siblings did appear to share components. In several instances, prolonged immunization of nonrestricted responders in these groups resulted in the production of structurally restricted antibodies. However, this was not a general phenomenon. Finally, some rabbits immunized with type III Pneumococcus produced restricted components that did not appear to possess anti‐polysaccharide activity.
Primary antibody formation in vitro by mouse cells in a complete homologous system
Authors: Munder P. G. , Modolell M. , Raetz W. , Luckenbach G. A.
Published: February 2007

Mouse lymphocytes have been cultivated in the absence of heterologous sera or mitogenic compounds. The fetal calf serum usually used was replaced by small amounts (0.5–1 mg protein/ml medium) of homologous exudate fluid induced by intraperitoneal injection of lysolecithin. Under these conditions, a primary immune response against various antigens (sheep red blood cells, dinitrophenyl‐dextran, dinitrophenyl‐human IgG) can be elicited. Adjuvant activity of lysolecithin has also been studied in this system.
Polymorphonuclear leukocyte stimulation by immune complexes. Assessment by nitroblue tetrazolium dye reduction
Authors: Nydegger U. E. , Anner R. M. , Gerebtzoff A. , Lambert P. H. , Miescher P. A.
Published: February 2007

Insoluble immune complexes and immune aggregates were found to stimulate the activity of the hexose monophosphate shunt as measured by nitroblue tetrazolium reduction in human polymorphonuclear leukocytes. This phenomenon depends more on the size of the complex than on its ability to activate complement.
Stimulation of DNA synthesis in mouse lymphoid cells by polyanions in vitro. II. Relationship between adjuvant activity and stimulation of DNA synthesis by polyanions
Authors: Vogt W. , Rühl H. , Wagner B. , Diamantstein T.
Published: February 2007

The effect of eleven different polyanions, among them dextran sulfate, and of two different preparations of dextran was investigated on [3H]dThd incorporation in spleen cells in vitro, and on PFC response to SRBC in vivo. In general, polyanions stimulated DNA synthesis in spleen cells and enhanced PFC response to SRBC.

Dextran itself had no effect on DNA synthesis and on PFC response. The degree of activation of DNA synthesis by polyanions in vitro was found to correlate with the adjuvant activity of the substances in vivo It is postulated that activation of DNA synthesis in lymphoid cells is essential for the adjuvant activity of polyanions.
Studies on immunological paralysis. XI. A comparison of the tolerogenicity in vitro of levan and type 3 pneumococcal polysaccharide
Authors: Kotlarski I. , Courtenay B. M. , Howard J. G.
Published: February 2007

Brief exposure of mouse spleen cell suspensions to levan (LE) in vitro induced a state of stable, specific unresponsiveness in these cells, as demonstrated by LE challenge 1 – 15 days after transfer of LE‐treated cells into lethally irradiated recipients. These mice showed unimpaired responsiveness to type 3 pneumococcal polysaccharide (SIII) and sheep erythrocytes as compared with recipients of untreated cells.

Suppression was attained by incubation in amounts of LE varying from 25 μg up to 10 mg/spleen equivalent and the increment in effect with rising concentrations of LE was relatively small. Tolerance induction in vitro was not dependent on the extremely high molecular weight of LE (mol. wt. 2 x 107) because it was equally demonstrable when hydrolyzed LE with a 1000‐fold reduction in average mol. wt. was used. Induction of unresponsiveness to LE could be prevented by exposure of spleen cells to anti‐mouse Fab serum prior to incubation with LE.

In contrast, attempts to produce similar suppression using SIII were completely unsuccessful. It is concluded that the difference in behavior of these two thymus‐independent antigens may be a reflection of weaker overall binding of SIII to IgM receptors on B cells than is the case with LE. Consistent with this view is the demonstration that highly depolymerized LE retained the capacity to inhibit the detection of LE‐specific plaque‐forming cells, whereas SIII of similar mol. wt. had lost this property.
Immune responses in vitro. IV. A comparison of the protein‐free and mouse serum‐supplemented mouse mixed lymphocyte interaction assays
Authors: Peck A. B. , Katz‐Heber Ellen , Click R. E.
Published: February 2007

Various parameters that optimize the cellular responses of mouse spleen lymphoid cells against alloantigens in a protein‐free and a mouse serum‐supplemented culture medium are defined. Results indicate: (a) The addition of sulfhydryl agents to either culture system caused an increase in thymidine incorporation, the percent increase being greater in allogeneic cultures than in syngeneic cultures. (b) The optimal ratio of responder cells to stimulator cells in the absence and presence of serum were different. The presence of mouse serum reduced by one‐half the number of stimulator cells necessary to maximize the response, while the optimal number of responder cells was similar in both systems. (c) The maximum incorporation of isotope for strain combinations which involved incompatibility at the major histocompatibility complex (H‐2) was at 66 h of culture in the protein‐free medium and at 98 h of culture in the serum‐supplemented medium.
Electrochemical reduction of immunoglobulins specific reduction of the inter‐heavy chain disulfide bridges of igg
Authors: Rivat C. , Fontaine M. , Ropartz C. , Caullet C.
Published: February 2007

Electrochemical reduction of IgG disulfide bridges has been studied and the number of disulfide bridges reduced determined by several methods: coulometry and polarographic titrations at the mercury electrode with electroactive alkylating agents, such as iodoacetamide and N‐ethylmaleimide. The three methods give identical results. Determination and localization of reduced disulfide bridges examined in the four IgG subclasses show that the reduction occurs exclusively on the interheavy chain disulfide bridges.
Cell‐mediated immune responses in vitro. v. a comparative study of in vitro immunogenicity of splenic lymphocytes, neoplastic lymphoid cells and fibroblasts
Authors: Wagner H. , Wyss C.
Published: February 2007

This study compares the effectiveness of mouse lymphocytes, neoplastic lymphoid cells or fibroblasts in stimulating allogeneic cells to embark on an in vitro cytotoxic response, as measured by a 51Cr release assay. In addition, during ontogeny of mouse spleen cells, their capacity to stimulate in the mixed lymphocyte culture (MLR) was compared to their capacity to stimulate cytotoxic allograft responses. During ontogeny, there was amarked increase in the capacity of mouse spleen cells to stimulate mitotic responses in the MLR. In contrast, the magnitude of cytotoxic allograft responses induced by neonatal mouse spleen cells in the cytotoxic allograft system was comparable in magnitude to that induced by spleen cells of adult mice.

The immunogenicity of subpopulations of mouse spleen cells was investigated. Mouse lymphoid cell populations, enriched for B or T lymphocytes or hemopoietic stem cells were equally immunogenic in vitro, as were myeloma or thymoma lymphoid cells. In contrast, mouse fibroblasts were found to elicit poor cytotoxic allograft responses. In fact, lymphoid cells were about 20–40‐fold more immunogenic than fibroblasts.
Specific priming of mouse thymus‐dependent lymphocytes to allogeneic cells in vitro
Authors: Andersson L. C. , Häyry P.
Published: February 2007

Blast cells from one‐way mixed lymphocyte cultures (MLC) revert to lymphocytes in vitro and in vivo. The blast‐derived lymphocytes (BDL) respond promptly in second MLC to the original and H‐2 cross‐reactive stimulator cells. The peak response by primed cells takes place on the 2nd – 3rd culture day, as compared to 6th – 7th day by nonprimed cells. BDL do not respond to the mitogens, phytohemagglutinin or concanavalin A.
Mitogenic properties of the thymus‐independent antigen pneumococcal polysaccharide siii
Authors: Coutinho A. , Möller G.
Published: February 2007

Pneumococcal polysaccharide type SIII was found to stimulate DNA synthesis in nonsensitized spleen cells cultivated in vitro, peak activation occurring 2 days after the addition of 150 μg/ml of SIII. Cortisone‐resistant thymocytes did not respond, whereas spleen cells from congenitally athymic (nude) mice reacted as normal spleen cells.

SIII induced polyclonal antibody synthesis in normal spleen cells, which was detected against a variety of red cell antigens and haptens. It was able to substitute for T cells, as demonstrated by the ability of sheep red cells to induce a primary immune response in cultures of nude spleen cells in the presence, but not in the absence, of SIII.

It is concluded that SIII is a B cell mitogen capable of directly activating DNA and antibody synthesis in these cells. This property is presumably responsible for the thymus‐independent nature of the immune response to SIII and lipopolysac‐charide. It is suggested that other thymus‐independent antigens also are B cell mitogens, although the degree of mitogenicity may vary.
The allogeneic effect: increased affinity of serum antibody produced during a secondary response
Authors: Elfenbein G. J. , Green I. , Paul W. E.
Published: February 2007

We have used the “allogeneic effect” as a model system for investigating the effects of augmented thymus‐derived lymphocyte function on secondary antibody responses of primed inbred guinea pigs. A transient graft‐versus‐host reaction was induced by infusing nonimmune strain 13 lymphoid cells into strain 2 guinea pigs which had been primed with dinitrophenylated (DNP) ovalbumin. Six days later the recipients were challenged with the priming antigen intradermally. Four days after boosting with antigen, sera from these “allogeneic” guinea pigs had significantly higher affinity for ε‐DNP‐L‐lysine than sera from “control” guinea pigs which were primed, but had not received allogeneic lymphoid cells. Spleen cells taken from “allogeneic” guinea pigs three to five days after boosting were shown to have both higher numbers of plaque‐forming cells and a predominance of high avidity antibody‐producing cells in comparison to spleen cells from “control” guinea pigs. Thus, increased donor thymus‐derived lymphocyte function brought about by the transient graft‐versus‐host reaction causes an enlargement of the host antibody‐producing cell pool and preferentially expands the subpopulation of cells producing high affinity antibody. The effects of this preferential expansion are reflected in the sera of “allogeneic” guinea pigs as higher affinity antibody in comparison to serum antibody of “control” guinea pigs.
Specificity of heterologous antisera to components solubilized from rat thymocyte membranes
Authors: Eshhar Z.
Published: February 2007

Rat thymocyte membranes were solubilized by citraconic anhydride and the soluble preparation was fractionated by gel chromatography. Antithymocyte sera that were elicited by injection of either these soluble components or intact thymocytes or membranes of thymocytes, were compared for their cytotoxic effect on various cell populations and prolongation of skin grafts. The different antisera were equally effective in cytolysis and lysed 100% of thymus cells, 60% and 15% of lymph node and bone marrow cells, respectively. The antiserum raised by the soluble components contained significantly fewer antibodies against red blood cells or kidney and liver tissues and was also much less toxic when injected into rats. The different antisera were also comparably effective in the prolongation of skin grafts. The soluble components of rat thymocyte membranes inhibited the cytotoxic activity of the antisera raised in rabbits by either cells or the soluble preparation. However, the soluble component did not inhibit the cytotoxic activity of alloantisera, suggesting that the soluble component does not contain active histocompatibility antigens.
The role of t cells in the passive transfer of contact sensitivity and their occurrence in the bone marrow
Authors: Zembala M. , Asherson G. L.
Published: February 2007

Contact sensitivity to 4‐ethoxymethylene‐2‐phenyl oxazolone can be transferred to irradiated recipients by lymph node, spleen and bone marrow cells from sensitized donors. The transfer by lymph node, spleen and bone marrow cells was abolished by anti‐θ serum whose specificity was confirmed by absorption with CBA brain. Transfer by peritoneal exudate cells was relatively resistent to treatment with anti‐θ serum. Mice deprived of T cells did not develop contact sensitivity following immunization, and their bone marrow and peritoneal exudate cells failed to transfer contact sensitivity. It was concluded that T cells were required both for the induction and for the passive transfer of contact sensitivity.
The allogeneic effect in vitro: activation of antibody synthesis in aggressor and target lymphocyte populations by allogeneic interaction
Authors: Möller G. , Coutinho A.
Published: February 2007

Normal F1 hybrid spleen cells, interacted in tissue culture with cortisone‐resistant thymocytes (CRT) of parental origin, were activated to increased antibody synthesis against SRC (sheep red cells) as compared to F1 hybrid lymphocytes mixed with syngeneic CRT. However, an equal or greater stimulation occurred when parental spleen cells were mixed with F1 hybrid CRT as compared to syngeneic mixtures. When the experiments were performed in cultures containing fetal calf serum, large doses of T cells suppressed the immune response to SRC in both directions. Supernatants from a mixture of parental CRT and normal F1 spleen cells stimulated the immune response of both parental and F1 spleen cells to SRC.

Analogous results were obtained when the secondary response was studied. Thus, spleen cells from mice hyperimmunized to SRC were activated to increased synthesis of indirect plaque‐forming cells by allogeneic confrontation. However, hyperimmune parental spleen cells were activated by confrontation with nonimmune F1 lymphocytes to the same extent or more than hyperimmune F1 spleen cells mixed with normal parental cells.

It is concluded that the stimulating effect on the immune response caused by allogeneic interaction does not exhibit specificity for the genotype of the target B lymphocytes. Possible mechanisms by which specificity can be detected by in vivo experiments are discussed.
The mechanism of antigenic competition. i. the macrophage as a site of a reversible block of t‐b lymphocyte collaboration
Authors: Schrader J. W. , Feldmann Marc
Published: February 2007

A model of antigenic competition, initiated in vivo, but expressed in vitro, was developed in order to study the cellular basis of antigenic competition. This was initiated by the injection of donkey or chicken red cells 2 days before sacrifice. To permit a response to a thymus‐dependent hapten protein conjugate in vitro, it was necessary to prime the mice with the carrier some weeks prior to culture. Thus the response of mice primed with fowl gamma globulin (FGG) to dinitrophenylated (DNP) FGG in vitro was used as the test for antigenic competition. The function of B cells was found to be normal, since the response to a thymus‐independent antigen, DNP flagella of Salmonella adelaide, was found to be normal, while that to DNP FGG was suppressed. The locus of suppression in antigenic competition was found to be the macrophage, since small numbers (3000) of peritoneal cells which had been depleted of T cells by treatment with anti‐θ serum and complement restored the response to DNP‐FGG. Trypsinization of the spleen populations also overcame antigenic competition, indicating that there was no irreversible block of any cell type. Trypsinization did not augment the in vitro response to DNP‐FGG of noninjected mice or that to DNP‐FLA of normal or DRC injected mice. Thus, trypsinization overcame competition without augmenting the response of either T or B cells and thus acted to remove a surface inhibitor from the surface of macrophages. The results were interpreted in the light of current concepts of the mechanism of T‐B collaboration. It is proposed that antigenic competition is a byproduct of the release of complexes of IgT and antigen by T cells. These bind to macrophages, initiating the response to the first antigen, but competing out the binding of IgT complexes to a subsequently administered antigen which are needed to trigger B cells and thus blocking the cooperative response to the second antigen.
Antibody isoelectric spectra visualized by antigen‐coated erythrocytes
Authors: Phillips Jennifer M. , Dresser D. W.
Published: February 2007

A method is described for visualizing isoelectric spectra of antibody with antigen‐coated erythrocytes. The method, which is quick and simple, can be used for a wide range of antigens. This hemolytic technique detects antibodies which are not detectable by the radiolabeling method.
Thymus reticulum cell cultures confer T cell properties on spleen cells from thymus‐deprived animals
Authors: Wekerle H. , Cohen I. R. , Feldman M.
Published: February 2007

The development of T lymphocyte immunocompetence appears to depend upon the function of the thymus gland. To study this function in greater detail, we developed a method for preparing monolayer cultures of thymus reticulum cells. We found that populations of lymphocytes from thymus‐deprived mice acquired the ability to produce T lymphocyte reactions when incubated with reticulum cells in vitro. These results provide evidence about the role of the thymus reticulum in inducing immunocompetence of T lymphocytes.
Transfer of low zone tolerance to normal syngeneic mice by Θ‐positive cells
Authors: Weber G. , Kölsch E.
Published: February 2007

Low zone tolerance (LZT) of mice to bacteriophage fd has been further investigated and found to be specific for the antigen and inducible within 24 h. LZT and immunity are retained when spleen cells from tolerant or immune animals are transferred into thymectomized, lethally irradiated recipients reconstituted with fetal liver. Thymocytes, educated in vivo with antigen doses inducing LZT, are capable of transferring this tolerance into normal syngeneic mice and maintaining it there specifically. Treatment with anti‐Θ serum abolishes the suppressive capacity of the educated thymocytes. The data suggest that LZT is actively maintained by living T cells.
Sequential recruitment of antibody class‐committed B lymphocytes during ontogeny
Authors: Ivanyi J.
Published: February 2007

Chickens were bursectomized surgically during the first week after hatching and immunized subsequently with human serum albumin (HSA). Bursectomy (Bx) 4 days posthatching suppressed the IgG response, but did not affect the IgM response when tested at 2 weeks. The overall responsiveness had declined by 4 weeks, however, probably due to the nonreplicating character of the early peripherally seeded B cells. Furthermore, these chickens failed to give a memory response. B cells with self‐replicating and memory potential are seeded later than 4 days posthatching. However, 7 day Bx still produced a selective suppression of the primary IgG response while the IgM response, although delayed, reached a normal peak titer. Treatment of memory cells in vitro with anti‐μ serum interfered with the adoptive transfer of both IgM and IgG responses. The data are interpreted in terms of sequential seeding of IgM and IgG precursors of the anti‐HSA clones, both of which maintain membrane‐bound μ‐chain antigen.
Immunoglobulin determinants on lymphocytes in germ‐free piglets
Authors: Jarošková Lydie , Trebichavský I. , Řiha I. , Kovářů F. , Holub M.
Published: February 2007

Surface IgM determinants on splenic lymphocytes were detected in 80‐day old pig fetuses using anti‐L and anti‐μ sera conjugated with horseradish peroxidase. Similarly, by the immunofluorsecence technique, only IgM determinants were demonstrated on unprimed lymphocytes from spleen and bone marrow in fetuses at the end of gestation (on the 112–114th day) and in precolostral, germ‐free piglets.

The pattern of labeling with anti‐μ sera is characteristic for nonactivated lymphocytes as documented on fetal splenic lymphocytes or the small lymphocytes in a dense lymphatic area of the omentum of precolostral piglets. The lymphocytes which showed surface immunofluorescence staining with anti‐γ serum were observed in conventionally reared, nonimmunized piglets and appeared in all the germ‐free reared piglets which were immunized with sheep red blood cells, human serum albumin or bacterial antigens.

We conclude that the true antigen recognition units, or “primary” receptors, on antigen‐sensitive uncomitted lymphocytes are exclusively of the IgM type and that the IgG‐carrying cells can be detected in piglets only after antigenic stimulation.
Antibodies specific for human T lymphocytes in cold agglutinin and lymphocytotoxic sera
Authors: Thomas D. B.
Published: February 2007

Sera of patients with cold agglutinin disease (anti‐I,i), systemic lupus erythematosus, infectious mononucleosis or cryoglobulinemia contained cold IgM antibodies cytotoxic for a subpopulation of normal lymphocytes. Evidence was obtained that the antibodies were specific for T lymphocytes. There was an enrichment of cells reacting with the sera after removal of Ig‐bearing cells on immuno adsorbent columns. Absorption studies with adult and cord erythrocytes revealed a minimum of three antigenic determinants on the lymphocyte membrane, viz. i,I and a third specificity which was unique to T cells. Antibodies to this third specificity were present, at high titer, in sera which were monoclonal for the i antigen.
Difference between the lymph node response to injection of autosensitized T lymphocytes and that in a graft‐versushost reaction
Authors: Cohen I. R.
Published: February 2007

Thymus (T) lymphocytes autosensitized in vitro were shown in previous studies to produce enlargement of draining popliteal lymph nodes upon injection into the footpads of syngeneic rats. Specific autoreactive effector lymphocytes were found to be recruited within these lymph nodes. In the present study, the cellular basis of lymph node enlargement in mice by autosensitized lymphocytes was compared with that produced in a graft‐vs.‐host (GvH) reaction. T lymphocytes of C3H mice were autosensitized against syngeneic fibroblasts in vitro for 16 to 18 h in the absence of serum, and 107 lymphocytes were injected into the footpads of syngeneic mice. Control lymphocytes were incubated without fibroblasts. The GvH reaction was produced by injecting 107 C3H T lymphocytes into the footpads of (C3H × C57BL)F1 adult recipients. The index of relative enlargement of the draining popliteal lymph nodes was measured 6 days after injection. Experiments were done to identify the origin of the lymph node cells in these reactions. Irradiation of the donor lymphocytes (1000 r) or the recipient mice (550 r) was used to prevent proliferation of the lymphocytes of either origin. The participation of recipient T lymphocytes in lymph node enlargement was investigated by using thymectomized mice.

The following results were obtained.

1) The GvH lymph node enlargement was found to depend on proliferation of the donor T lymphocytes, but did not seem to require the participation of radiosensitive cells within the recipient mice.

2) In contrast, the response of the lymph nodes to autosensitized donor T lymphocytes depended on the function of radiosensitive T lymphocytes within the syngeneic recipients. The autosensitized donor lymphocytes themselves did not have to proliferate to recruit the response of recipient T lymphocytes.

3) It was found that recruitment of recipient lymph node cells could be super‐imposed upon a conventional GvH reaction by presensitizing the C3H donor lymphocytes in vitro. Both autosensitization against syngeneic or allosensitization against C57BL fibroblasts augmented the lymph node response of (C3H × C57BL)F1 hybrid recipients. The recruited or donor components of these mixed responses could be selectively abolished by irradiating either the donor lymphocytes or the recipient mice.

Hence, the autosensitization response, like the host‐vs.‐graft transplantation reaction, can be induced by sensitization of lymphocytes peripherally and involves recruitment of lymphocytes within regional lymph nodes. The GvH response manifested in the same popliteal lymph nodes does not appear to require the recruitment of radiosensitive T lymphocytes.

These findings suggest that different classes of T lymphocytes function in the autosensitization and GvH responses.
Synthesis and immunological properties of an artificial antigen with the repeating oligosaccharide unit of Salmonella illinois as haptenic group
Authors: Kleinhammer G. , Himmelspach K. , Westphal O.
Published: February 2007

The serological specificity of Salmonellae (O‐specificity) is mediated by oligosaccharide subunits (repeating units) of the species‐specific cell wall polysaccharides. With S. illinois (O‐specificities 3, 15 and 34 of the Kauffmann‐White scheme) as a representative example, it was possible to achieve the coupling of a complete repeating unit of that kind (in the case of S. illinois a tetrasaccharide) to protein, thus converting it into an artificial antigen. The repeating tetrasaccharide of the O‐specific S. illinois polysaccharide has been obtained by partial acid hydrolysis of the O‐specific S. illinois lipopolysaccharide and has the following structure with rhamnose as the reducing terminal sugar: magnified image In a reaction of this tetrasaccharide (TSill) with o‐phenylenediamine and m‐nitrophenylhydrazine and its subsequent reduction, TSill‐illl‐(3‐amino‐phenyl)‐flavazole was prepared, which was then coupled to edestin by means of the azo‐method.

Antisera obtained after immunization of rabbits with the TSill‐illl‐(3‐azophenyl)‐flavazole‐edestin conjugate contained TSill‐specific antibodies, as shown by agglutination of S. illinois bacteria as test antigen (titers up to 1:2500). The specificity of these antibodies was tested in detail by cross‐agglutination with various bacterial strains. These antibodies are predominantly directed against the non‐reducing end group Glc magnified image Gal magnified image (Man) (O‐factor 34), and only to a slight extent against the internal groupings' Gal magnified image Man (O‐factor 15) and Man magnified image (Rha) (O‐factor 3). The serum antibodies did not show cross‐reactions with O‐factor 122, which differs from O‐factor 34 in the configuration of the Galmagnified imageMan‐linkage (122 :α‐glycosidic, 34:β‐glycosidic). Thus, by immunization of rabbits with a TSill‐illl‐(3‐azophenyl)‐flavazole edestin conjugate, a largely factor‐specific (anti‐34) antiserum was obtained.
Macrophages bearing hapten‐carrier molecules as foci inducers for T and B lymphocyte interaction
Authors: Askonas Brigitte A. , Roelants G. E.
Published: February 2007

In a syngeneic cell transfer system in C3H/He mice, we studied the effect of antigen‐bearing macrophages on the expression of B memory cells specific for the 2,4‐dinitrophenyl (DNP) group. The development of anti‐DNP clones was enhanced by macrophages carrying antigen on transfer of limiting dilutions of DNP‐BGG (bovine IgG)‐primed spleen cells. Triggering of B memory cells in irradiated hosts was enhanced both in terms of clonal expression with resulting high levels of antibody production, and of the number of anti‐DNP clones (B memory cells) colonizing the spleen effectively.

To observe this enhancement, both hapten and carrier determinants needed to be carried by the same macrophage. The results are most easily interpretable by assuming that antigen‐carrying macrophages provide a center for the attraction of T and B cells in a spleen focus where they can interact more efficiently.
Anti‐lymphocytic antibodies and marrow transplantation. III. Effect of heterologous anti‐brain antibodies on acute secondary disease in mice
Authors: Rodt H. , Thierfelder S. , Eulitz M.
Published: February 2007

Antisera against murine thymus‐derived lymphocytes were raised in rabbits injected with mouse brain. It was possible to remove contaminating antibodies which cross‐reacted with B cells and hemopoietic stem cells by absorption with liver and plasmacytoma followed by purification and fractionation over DEAE‐cellulose. T cell specificity of the absorbed anti‐mouse brain serum (AMBS) and its purified globulin fraction (AMBG) was demonstrated by cytotoxicity, complement fixation and unaltered numbers of plaque‐forming cells and colony‐forming units. In vivo, the effect of absorbed AMBG on the graft‐versus‐host reaction was investigated in an H‐2‐incompatible parent‐to‐F1 hybrid donor‐recipient combination. (C57BL/6 × CBA)F1 mice irradiated with 900 rad and transfused with C57BL/6 spleen cells died from acute secondary disease within 20 days. They survived as complete chimaeras beyond day 300 without symptoms of secondary disease when the donor cells had been preincubated with absorbed AMBG. The implications of the suppression of secondary disease in this model are discussed.
The use of protein A‐containing Staphylococcus aureus as a solid phase anti‐IgG reagent in radioimmunoassays as exemplified in the quantitation of α‐fetoprotein in normal human adult serum
Authors: Jonsson S. , Kronvall G.
Published: February 2007

A new technique for the separation of antigen‐antibody complexes from free antigen in radioimmunoassays has been developed. The technique is based on the interaction between the Fc portion of IgG and staphylococcal protein A. One ml of a 10 % suspension of formaldehyde‐treated and heat‐killed staphylococci of strain Cowan I, particularly rich in protein A, at equivalence binds 1.2–1.5 mg IgG in a pure IgG solution and 0.8–1.0 mg of IgG present in serum. This staphylococcal solid phase anti‐IgG reagent is added in excess of the IgG content in the preincubated radioimmunoassay reaction mixture. Centrifugation can be started within minutes. The bacteria form a solid pellet, so the supernatant can be decanted without further washing being required. Nonspecific uptake of radioactivity is below 7 %. In our assay for human α‐fetoprotein using antiserum raised in the rabbit, completeness of separation is better than 98 %. The present technique will differentiate between the presence of antigen and homologous IgG antibody in a sample tested.
Synthesis of the second and fourth components of complement by tissue culture cell lines
Authors: Wyatt H. V.
Published: February 2007

A number of tissue culture cell lines were tested for synthesis of C1, C2 and C4. Supernates from two human embryo cell lines were positive for C1 when received, but on further culturing gave only slight amounts of C1. A number of cell lines produced C2 over a long period. This production was followed in hybrid cells formed by cell fusion. Some mixed rat liver cell cultures produced both C2 and C4. Of 30 tissue cultures tested, seven were found to synthesize C2.

A cell line derived from guinea pig peritoneal cells produced C2 when grown with heat‐inactivated serum or in a synthetic medium. About 310 effective C2 molecules were produced per hour per cell. Unlike fresh peritoneal cells, production of C2 by the cell line was unaffected by hydrocortisone. Addition of stimulated or unstimulated lymphocytes with or without macrophages did not alter C2 production.
Production of anti‐H‐2 antibodies in thymectomized mice
Authors: Klein J. , Livnat S. , Hauptfeld V. , Jeřábek L. , Weissman I.
Published: February 2007

Neonatally and adult thymectomized (Tx) mice of strains C57BL/10Sn and A. BY (H‐2b) were immunized at the age of 5 – 8 weeks with lethally irradiated spleen and lymph node cells from adult B 10. A and A (H‐2a) donors, respectively. The titer of H‐2 hemagglutinins and cytotoxins was determined at different intervals after immunization. Both untreated antibodies and antibodies reduced with 2‐mercaptoethanol (2‐Me) were tested. The non‐Tx control mice responded with production of both 2‐Me‐sensitive and 2‐Me‐resistant H‐2 antibodies. In the Tx mice, only 2‐Me‐sensitive H‐2 antibodies were detected. Thus, the anti‐H‐2 IgM response appears to be thymus‐independent and the anti‐H‐2 IgG response thymus‐dependent.
Three classes and four (sub)classes of rat immunoglobulins: IgM, IgA, Ige and IgG1, IgG2a, IgG2b, IgG2c
Authors: Bazin H. , Beckers A. , Querinjean P.
Published: February 2007

Class or subsclass‐specific differences have been investigated in the monoclonal immunoglobulins produced by 184 secreting immunocytomas raised in the LOU/Wsl inbred strain of rats. Seven classes or subclasses of rat immunoglobulins were detected and the names IgM, IgA, IgE, IgG1, IgG2a, IgG2b and IgG2c proposed for them. The sedimentation coefficients of rat IgG1, IgG2a and IgG2c were found to be respectively, 6.7, 6.4 and 6.7 S. Rat IgG1, IgG2a and IgG2c were found to cross‐react with antisera to human γ‐chains.
Hapten inhibition of the immune response in vitro
Authors: Kettman J.
Published: February 2007

The in vitro primary immune response of mouse spleen cells against a hapten, the 2,4,6‐trinitrophenyl (TNP) group, bound to an erythrocyte carrier is inhibited by the polyvalent, but nonimmunogenic TNP17HSA (human serum albumin) when added at the same time as the immunogen. Resistance to the effect of TNP17HSA develops in 24 h after the addition of immunogen. Sensitivity to this inhibition can be quickly re‐established after a mild physical dissociation of cell aggregates in the spleen cell cultures even 48 h after the start of a response. This sensitivity is rapidly lost again if the culture are incubated for a few hours before the TNP‐HSA is added.
Cooperation by mouse T lymphocytes: the role of antibody in T cell specificity
Authors: Playfair J. H. L. , Marshall‐Clarke S. , Hudson L.
Published: February 2007

Mouse thymus cells were educated in vivo with horse red blood cells (RBC) and tested for their ability to cooperate in the response of bone marrow cells to sheep RBC. Significant cooperation was observed only when anti‐sheep serum was injected into the educating mouse. We suggest that cooperation and some other properties of the T cell are mediated by passively acquired antibody.
Familial occurrence of isolated IgA deficiency associated with antibodies to IgA. Evidence against a structural gene defect
Authors: van Loghem Erna
Published: February 2007

Two families were investigated for the occurrence of IgA deficiency and antibodies to IgA. Several members of the families were found to be deficient for IgA1, as well as for IgA2, which strongly suggested that the disorder is genetically determined. By means of typing for genetic markers a defect of structural α genes could be excluded.

An autosomal recessive mode of inheritance of genes committed with the synthesis of IgA accounts best for the observations.

Anti‐IgA antibodies of different specificity were encountered in eight out of fifteen IgA‐deficient family members; sometimes more than one specificity was observed in the same individual. There was no association with Gm allotypes, nor any evidence for a possible stimulation of antibody production.
Selective activation of mouse T and B lymphocytes by periodate, galactose oxidase and soybean agglutinin
Authors: Novogrodsky A.
Published: February 2007

Modifications of surface saccharides of mouse spleen cells wirh periodate (NaIO4) or with neuraminidase and galactose oxidase induced extensive blastogenesis. The mitogenic effects of these agents on purified thymus‐derived lymphocytes (T cells) and bone marrow‐derived lymphocytes (B cells) were investigated. Thymus cells were stimulated after treatment with NaIO4 or with neuraminidase and galactose oxidase. The stimulation of these cells was markedly enhanced upon addition of a lymphocyte‐activating factor obtained from human leukocyte culture. Spleen cells from congenitally athymic mice did not respond to treatment with NaIO4 or with neuraminidase and galactose oxidase. Spleen cells from mice deprived of T cells by thymectomy, irradiation and bone marrow reconstitution, responded poorly to the above mentioned agents. It was concluded that periodate or treatment with neuraminidase and galactose oxidase stimulates exclusively T cells. Soybean agglutinin (SBA) had selective effects on T and B mouse lymphocytes. Neuraminidase treatment rendered T cells highly responsive to stimulation by SBA. On the other hand the small but significant stimulation of B cells by SBA is reduced after treatment of the cells with neuraminidase. SBA stimulates B cells and neuraminidase‐treated T cells by interaction with galactose‐like sites.