bluuebunny/crossref_metadata_2025 huggingface dataset:
Do Ly Phenotypes Correlate with Function or with MHC Determinant Recognized?
Authors: Swain Susan L. , Panfili Peter R. , Dutton Richard W.
Published: December 2022

Many observations have led to a general hypothesis in which T cells that mediate helper functions have the Ly phenotype 1+,2- and are specific for I region determinants whereas T cells which mediate suppressor functions are Ly1+ or -,2+ and preferentially recognize K or D determinants. However, there are some exceptions to this dichotomy. We have shown that allogeneic helper effects can be generated to K and D determinants and suppression can result from I region differences. These exceptions raise the question of which factor determines the Ly phenotype of the T cell—the function of the cell or the MHC determinant that is recognized by the cell. In the experiments to be presented, we show that those helper T cells stimulated by K, D differences are Ly2+, while those stimulated by I region differences are Ly2-.

Positive allogeneic effects were determined by titrating mitomycin-treated, nylon wool passed splenic T cells into allogeneic B cells (anti-T treated spleen) plus SRBC.
Clones of Alloreactive T Cells
Authors: Fathman C. G. , Hengartner H.
Published: December 2022

Previous observations from our lab and many others have suggested that MLR determinants are composed of I region gene products. We extended our original observations by studying the reactivities of responder cells obtained after in vitro primary stimulation (primed responder cells = PRC) with selected stimulator cells. These studies suggested that MLR determinants exist as “interaction” products of several I region genes which can act in cis- or trans- position. Furthermore, these studies suggested the existence of specific clones of PRC which could recognize unique F1 MLR determinants. Using serially restimulated PRC which had been in culture for over three months, we derived colonies of alloreactive cells (using soft agar cloning techniques) which have remarkably restricted reactivities. Studies with these “cloned” PRC supported our original premise concerning clones of unique F1 reactive PRC and suggested the existence of unique “homozygous” MLR determinants and clones of cells which specifically recognize such determinants.
Characterization of Ia Antigens on Trinitrophenyl (TNP)-Modified Guinea Pig Macrophages
Authors: Clement L. T. , Thomas D. W. , Shevach E. M.
Published: December 2022

The proliferative response of hapten-reactive guinea pig T lymphocytes to TNP-modified macrophages is regulated by the I-region of the MHC of the priming macrophage and can be specifically inhibited by both anti-TNP serum and anti-Ia serum directed against the stimulator macrophage. Although these findings suggest an intimate association of macrophage Ia antigens and the TNP-specific immunogen, the precise nature of the antigenic determinant(s) recognized by the T cell remains unclear. We have investigated this apparent association by chemical methods. Peritoneal exudate cells from strain 13 guinea pigs were radio-labeled with 125Iodine or 3H-leucine, derivatized with trinitrobenzene sulfonate, and lysed with detergent. TNP-modified proteins were then precipitated with anti-TNP serum and analyzed by SDS-polyacrylamide gel electrophoresis. Although multiple membrane proteins were precipitated by anti-TNP serum, sequential precipitation experiments using anti-TNP or anti-Ia sera demonstrated that the TNP-bearing molecules and Ia antigens were distinct and separate from one another.
Macrophage-Lymphocyte Interactions Regulated by the MHC
Authors: Unanue Emil R. , Farr Andrew G. , Beller David I.
Published: December 2022

Macrophages can regulate lymphocyte proliferation and differentiation by various mechanisms, some of which are under the control of the MHC gene complex. Three distinct functions of the macrophage can be identified: 1) antigen presentation; 2) stimulation or inhibition of lymphocyte proliferation, resulting from secretion of mitogens or inhibitors; this function is modulated by phagocytosis of antigen, interaction with lymphocytes, and the state of macrophage activation; and 3) control of antigen-independent differentiation, as it takes place, for example, during part of thymocyte maturation.

Antigen presentation can be regulated by the MHC gene complex in an in vitro culture system involving a brief interaction of macrophages that have phagocytized Listeria monocytogenes and immune T cells. Such cultures contain copious amounts of a thymocyte mitogen, the secretion of which requires homology at the I-A region between the T cell and the macrophage and intimate contact for at least six hours.
Synergism Between Two Types of I-Region-Restricted Helper T Cells
Authors: Swierkosz James , Marrack Philippa , Kappler John W.
Published: December 2022

In vitro interactions between KLH, SRBC, or (T,G)-A--L-specific helper T cells and either B cells or macrophages (Mϕ) were shown to involve the simultaneous recognition of both the antigen and a product(s) of the I-region expressed on the B cell and the Mϕ. For example, in the anti-TNP response to TNP-(T,G)-A--L, T cells from (T,G)-A--L primed high × low responder F1 mice required that both high responder B cells and Mϕ be present in culture for a high response. Thus, it was surprising that KLH or SRBC primed T cells from F1 mice whose parents differed at H-2 could interact equally well with Mϕ-depleted B cells from either parent, regardless which parental Mϕ was used to reconstitute the cultures.

One possible explanation for these results was that effective responses required two synergising antigen-specific, I-specific helper T cells, one of which interacted with antigen bound to the Mϕ, and the other of which interacted with antigen bound to the B cell.
H-2 Gene Control of T-B Collaboration in Vivo
Authors: Sprent Jonathan
Published: December 2022

Experiments on the response to sheep erythrocytes (SRC) in vivo indicate that H-2 genes control (i) the induction of T helper (Th) cells (presumed to be a reflection of T-“macrophage” interactions) and (ii) T-B collaboration. Studies on the helper function of normal and chimeric T cells activated to SRC in irradiated syngeneic and allogeneic mice suggest that antigen presentation by “macrophages” of a particular H-2 haplotype generates a population of activated Th cells which collaborate only with specific B cells carrying the same H-2 determinants as the “macrophages”. From this approach it has been concluded that in terms of their helper function (i) normal homozygous T cells of strain a consist of a single population of “anti-self a” (anti-a) Th cells which recognize antigen presented by “macrophages” or specific B cells of strain a but not by strain b cells.
Lack of Strain Restrictions Among Allogeneic Nonresponder Donors and Recipients of GAT-Specific T Cell-Derived Suppressor Factors
Authors: Kapp Judith A.
Published: December 2022

The synthetic terpolymer of l-glutamic acid60-l-alanine30-l-tyrosine10 (GAT) fails to stimulate development of GAT-specific antibody responses in nonresponder mice but stimulates development of GAT-specific suppressor T cells that inhibit the development of normal anti-GAT PFC responses to GAT complexed to methylated bovine serum albumin. Extracts from lymphoid cells of GAT-primed but not control, nonresponder (DBA/1) mice contain a T cell factor (GAT-TsF) that also specifically suppresses responses to GAT-MBSA by normal syngeneic spleen cells. The experiments to be reported demonstrate that: 1) extracts from all GAT-primed nonresponder mice tested contain (GAT-TsF; 2) non-H-2 genes do not restrict the production of GAT-TsF; 3) all nonresponder strains of mice regardless of their non-H-2 genes are suppressed by GAT-TsF from all other strains bearing the nonresponder H-2p,q,s haplotypes; 4) both syngeneic and allogeneic nonresponder mice are suppressed by purified GAT-TSF that lacks immunoreactive GAT; and 5) suppression of GAT-MBSA responses by both syngeneic and allogeneic nonresponder spleen cells is mediated by a molecule encoded by the H-2 gene complex.
Localization of Hepatitis A Antigen in Liver Tissue by Peroxidase-Conjugated Antibody Method: Light and Electron Microscopic Studies
Authors: Shimizu Yohko K. , Mathiesen Lars R. , Lorenz Douglas , Drucker Jacques , Feinstone Stephen M. , Wagner Jo Ann , Purcell Robert H.
Published: December 2022

Hepatitis A antigen (HAAg) was localized in liver tissue from marmosets inoculated with human hepatitis A virus (HAV) by light and electron microscopy by using a peroxidase-conjugated antibody method. The fine granular peroxidase staining was scattered throughout the cytoplasm of liver cells when viewed with the light microscope. The distribution of HAAg-positive cells was focal. Virus-like particles, 24 to 27 nm in diameter, were observed in the cytoplasm of hepatocytes and smaller cells, resembling Kupffer cells, by standard thin-section electron microscopy (thin section EM). By immunoperoxidase electron microscopy (immunoperoxidase EM), HAAg was detected on the particles, which were aggregated within cytoplasmic vesicles of the hepatocyte. The surrounding membrane of the vesicles was also HAAg-positive. Similar HAAg particles were observed in the cytoplasm of smaller cells adjacent to hepatocytes as well. Thus, immunoperoxidase EM revealed that the 24- to 27-nm virus-like particles in the cytoplasma of liver cells obtained from marmosets were infected with HAV contained HAAg.
p30-Anti-p30 Immune Complexes: Intravascular Clearance and Extravascular Sequestration in Rats Bearing Moloney Sarcoma
Authors: Charles Jennette J. , Feldman J. D.
Published: December 2022

Complexes of 125I p30, a viral core polypeptide, and rat anti-p30 antibody, preformed in vitro, were injected into the heart of BN rats bearing Moloney sarcomas (MST) and of BN rats bearing an unrelated tumor or unexposed to tumor. Complexes were cleared from the circulation of MST-bearing rats more rapidly than from sera of controls and were almost completely eliminated after 30 hr. There was no relationship between rate of disappearance and size of tumor or levels of circulating complexes. Disappearance rates in rats with progressing and regressing tumors were similar. Uncomplexed labeled p30 was cleared from the circulation of tumor-bearing and control rats with kinetics similar to those of labeled complexes. Complexes were localized in the spleen of tumor-bearing and control rats, but much more in spleens of MST-bearing rats. No other tissues, including tumor, concentrated complexes, nor was there binding to peripheral blood and spleen cells. The data suggest that augmented clearance and sequestration were due to the formation of large insoluble complexes that were rapidly removed by the reticulo-endothelial system.
Hapten-Specific T Lymphocyte Activation by Glutaraldehyde-Treated Macrophages: An Argument against Antigen Processing by Macrophages
Authors: Thomas David W.
Published: December 2022

In this communication the effects of glutaraldehyde treatment of trinitrophenyl-(TNP) modified macrophages on their ability to stimulate TNP-specific guinea pig T lymphocyte proliferation were studied. TNP-modified macrophages briefly treated with glutaraldehyde retained much of their ability to stimulate TNP-primed T cells. In contrast, similar treatment of allogeneic macrophages or soluble protein antigen-pulsed syngeneic macrophages completely eliminated their ability to stimulate a mixed leukocyte reaction or protein antigen-specific proliferation, respectively. TNP-modification did not appear to interfere with glutaraldehyde reactivity since macrophages treated with glutaraldehyde before or after TNP-modification stimulated equivalent T cell responses. However, glutaraldehyde treatment of TNP-modified macrophages that had been cultured overnight dramatically reduced their ability to stimulate TNP-specific T cells. Glutaraldehyde-treated TNP-modified macrophages also expressed the same genetic restrictions of T cell activation as untreated stimulators. Thus, T cells primed with syngeneic TNP-modified macrophages were restimulated only by glutaraldehyde-treated TNP-modified syngeneic, but not by allogeneic, macrophages. These results are discussed with respect to the nature of the TNP-specific immunogen recognized by T cells.
Natural Cytotoxic Cells against Solid Tumors in Mice
Authors: Paige Christopher J. , Figarella Elizabeth Feo , Cuttito Michael J. , Cahan Anthony , Stutman Osias
Published: December 2022

The following characteristics of the effector cells for natural cytotoxicity (NC) against BALB/c Meth A tumor cells in BALB/c and other mouse strains were observed: 1) time course studies showed that most of NC was completed by 12 to 24 hr of incubation; 2) pre-incubation at 37°C for as long as 6 days had no effect on NC activity; 3) pre-treatment with ammonium chloride had no effect on NC activity; 4) NC cells were not adherent to plastic surfaces; 5) NC cells showed some degree of adherence to nylon wool fibers and activity was recovered from the adherent population; 6) NC cells were partially retained by G10-Sephadex columns; 7) by using velocity sedimentation at unit gravity in Ficoll gradients, NC cells were homogeneous having an average sedimentation velocity of 4.3 to 4.5 mm/hr; 8) treatment with carbonyl iron and magnetism had no effect on NC activity; 9) treatment with various antisera + C including, anti-Thy 1, anti-Lyt 1, anti-Lyt 2, anti-Ala 1, anti-NK, rabbit anti-mouse brain, and rabbit anti-mouse Ig, had no effect on NC activity; 10) heat-aggregated Ig had no effect on NC-mediated CMC; 11) trypsin treatment decreased NC activity but the effect was reversible, dependent on trypsin dose, and detected only in short-term CMC assay. These properties were similar for NC cells obtained from normal or nude mice, as well as peritoneal exudates obtained 5 days after i.p. BCG. When compared to the properties of the NK cell, many similarities were evident although differences in sensitivity to pre-incubation at 37°C and in the relative degree of adherence to nylon fibers were found. It is proposed that both the NC and NK cells belong to a family of effector cells with similar functions.
Cell-Mediated Destruction of Cells Grown on Artificial Capillaries
Authors: Zwilling Bruce S. , Clayman David A.
Published: December 2022

This investigation was designed to determine the conditions required to assess cell-mediated destruction of target L-cells grown on artificial capillaries. In control cultures that contained L-cells alone, solid nodules with a diameter of 1 mm as well as dense cellular growth could be visually observed by the 12th day of culture. Alloimmune spleen cells from both immunized and normal C57BL/10 mice were shown to be capable of destroying tritiated thymidine-labeled L-cells growing on artificial capillaries. The destruction of target cells grown as monolayers in capillary culture correlated well with monolayer cultures incubated in 16-mm plastic tissue culture wells. When target cells were grown in capillary culture for 5 days before the addition of effector cells, significant destruction by normal effector cells was not observed until the 15th day of culture whereas that mediated by immune cells was observed by the 7th day. The possible effects of cell-culturing conditions on the kinetics of cell-medicated destruction in capillary chambers are discussed.
Serologic Typing of Rickettsiae of the Spotted Fever Group by Microimmunofluorescence
Authors: Philip Robert N. , Casper Elizabeth A. , Burgdorfer Willy , Gerloff Robert K. , Hughes Lyndahl E. , John Bell E.
Published: December 2022

The micro-immunofluorescence (micro-IF) method was used to type rickettsiae belonging to the spotted fever or typhus groups according to their surface antigens. Seventy-two strains of rickettsiae from diverse sources, with varying histories of laboratory manipulation and immunologic characterization by other methods, were cross-tested against their mouse antisera. Fifteen serologic patterns (serotypes) were observed, 12 associated with spotted fever-group (SFG) rickettsiae and three with typhus group (TG) organisms. The reaction patterns of strains within each serotype were homogeneous. Nine of the SFG serotypes and the three TG serotypes were characteristic for rickettsiae that have been classified by other biologic procedures as to species (classified serotypes). The classified SFG serotypes included Rickettsia rickettsii (R-like), R. rickettsii (Hlp-like), R. sibirica, R. parkeri, R. conorii, R. rhipicephali, R. montana, R. australis, and R. akari. The three TG serotypes included R. prowazekii, R. typhi, and R. canada. Three SFG serotypes included rickettsiae that have not been identified as to species. Each consisted of two or more strains isolated in different years from widely-spaced localities in the United States and were considered to be distinct serotypes, which for the time being are unclassified. They included two strains of rickettsiae isolated from Dermacentor occidentalis ticks in California, three strains recovered from D. parumapertus ticks in southwestern United States, and three strains obtained from Ixodes pacificus ticks in Oregon. Each of three other strains had patterns of reaction that differed from those of all other rickettsiae. However, they were single representatives and evidence derived from this study is not sufficient to consider these as comprising additional serotypes.

The results obtained by micro-IF are in general agreement with other procedures for antigenic differentiation of rickettsiae belonging to the SFG and TG. Therefore, an immunologic basis for serologic classification of these rickettsiae is likely. Micro-IF should prove to be particularly useful for determining taxonomic and epidemiologic relationships among SFG rickettsiae because of its simplicity and general applicability.
Lack of Generation of Killer Cells in the Mixed Lymphocyte Reaction Between Mitogen-Stimulated and Unstimulated Autologous Lymphocytes
Authors: Katz Paul , Haynes Barton F. , Fauci Anthony S.
Published: December 2022

The present study has demonstrated that the Con A-activated cell-mediated autologous mixed lymphocyte reaction (MLR) is not associated with the generation of cytotoxic effector cells that kill autologous targets. Thus, the suppression of antibody production of PWM stimulated lymphocytes by autologous Con A-activated suppressor cells cannot be explained by detectable cytotoxicity. We have further demonstrated that the stimulator cell in this system is a nonadherent non-T cell.
Studies on Immunosuppression by Cobra Venom Factor
Authors: Martinelli Giorgio P. , Matsuda Takemasa , Waks Harriet S. , Osler Abraham G.
Published: December 2022

CVF administered before immunization can profoundly depress humoral responses in C-sufficient mice. In AKR/J C5- mice given CVF before i.v. immunization with SRBC, only IgG levels were depressed, IgM titers being equivalent to those of untreated controls. The immunosuppressive effect became inapparent when the i.p. route of immunization was adopted. In DBA/2J C5- mice reduction of both IgG and IgM titers was observed irrespective of the route of immunization. The degree of suppression was, however, much more marked when mice were challenged intravenously. Essentially identical results were obtained with C5+ DBA/1J mice. These studies indicate that immunosuppression by CVF is unrelated to activation of the late C components. The significance of these findings is discussed with reference to the possibility that the generation of biologically active C fragments in conjunction with a C3 deficiency may account for immunosuppression by CVF.
Restricted Complement Activation by Escherichia Coli with the K-1 Capsular Serotype: A Possible Role in Pathogenicity
Authors: Stevens Paul , Huang Shena N-Y. , Welch William D. , Young Lowell S.
Published: December 2022

Eighty per cent of blood culture isolates of K-1 Escherichia coli are “resistant” to in vitro opsonization and phagocytosis by normal human leukocytes and serum. To investigate why most of bacteremic K-1 E. coli are poorly opsonized by normal human serum, we undertook this study to assess the serum opsonic requirements for K-1 isolates particularly in regard to the role of the alternative pathway of complement activation. To ascertain the role of complement in opsonization, we used an in vitro opsonic and bactericidal killing system and a method that determines complement activation by measuring consumption of hemolytic complement. For both assays, MgEGTA was used to specifically block the classic pathway while leaving the alternative intact. Additionally, we used the bactericidal killing assay to determine the possible participation of specific opsonins, like antibody, in the opsonization of these E. coli by using a serum source that was either previously absorbed with bacteria to remove antibody or by using purified K-1 capsular antigen to block specific K-1 antibody. Data demonstrate that E. coli K-1 are opsonized only by the classic pathway of complement via antibody that is not specific for the capsular K-1 antigen. Furthermore, the hemolytic complement consumption method was shown to be far less sensitive in determining which complement pathway is activated during opsonization of E. coli K-1. The restriction of these bacteremic K-1 E. coli isolates to the classic pathway of complement activation and the requirements for specific antibody for effective opsonization may be important factors underlying the pathogenicity of these bacteria.
Cell Surface Glycoproteins of Rat Lymphocytes
Authors: Mitchell Richard N. , Bowers William E.
Published: December 2022

Oxidation of viable rat lymph node lymphocytes with either periodate or a combination of neuraminidase and galactose oxidase (NGO), followed by reduction with tritiated sodium borohydride, labels similar sets of cell-surface molecules as assessed by sodium dodecyl sulfate-polyacrylamide slab gel electrophoresis. Periodate and NGO induce blast transformation of lymph node lymphocytes (oxidative mitogenesis), and borohydride reduction inhibits the proliferative response. Thus, it is inferred that some or all of the glycoproteins that are labeled with tritiated borohydride may be involved in mediating the stimulation caused by the oxidizing agents.

Treatment of lymph node lymphocytes with 5 units/ml papain abolishes the response to periodate or NGO but does not significantly affect the response to Con A. At the same time, papain treatment eliminates the labeled bands representing six high m.w. glycoproteins (175,000, 170,000, 160,000, 155,000, 100,000, and 70,000 daltons). No significant effect is seen on the labeling of the other components visualized in the slab gels. The results implicate the subset of six high m.w. papain-sensitive sialoglycoproteins in mediating oxidative mitogenesis of rat lymph node lymphocytes.
Tumor Neutralization, Immunotherapy, and Chemoimmunotherapy of a Friend Leukemia with Cells Secondarily Sensitized in Vitro:
Authors: Cheever Martin A. , Greenberg Philip D. , Fefer Alexander
Published: December 2022

The aim of this study was to determine if cells primed in vivo could be rendered more effective in killing tumor by secondary sensitization in vitro. C57BL/6 mice were primed with a viable antigenic syngeneic Friend leukemia (FBL-3). The immune spleen cells secondarily sensitized by 5-day culture with irradiated FBL-3 (CiFx) became significantly more effective than fresh noncultured immune cells (Ci) in in vitro cytotoxicity, tumor neutralization (Winn assay), and immunotherapy of an early leukemia. However, in chemoimmunotherapy of advanced leukemia, CiFx were no more effective than Ci. Moreover, Ci cultured with irradiated C57BL/6 spleen cells (CiCx), i.e., without secondary sensitization in vitro, became markedly less effective than noncultured Ci. C57BL/6 mice inoculated with 5 × 106 FBL-3 on day 0 received on day 5 cyclophosphamide (CY) plus Ci, CiFx, or CiCx. CY alone doubled the median survival time (MST) to day 26. As an adjunct to CY, Ci or CiFx prolonged the MST to day 53 and day 55 and cured 16/72 and 20/66 mice, respectively, whereas CiCx prolonged the MST to only day 33 and cured 1/36 mice. To determine if the lack of enhanced effect after secondary sensitization in vitro reflects the nature of primary exposure to antigen, other priming regimens were studied. When mice were primed with irradiated FBL-3 or with cross-reacting MSV, CiFx were more effective than Ci in chemoimmunotherapy. Thus, enhancement by secondary in vitro sensitization of primed cells to eradicate advanced leukemia is dependent upon the method of priming and cannot be predicted by results of the in vitro tumor cytotoxicity, tumor neutralization, or immunotherapy of early leukemia. Moreover, primed cells lose effectiveness in tumor therapy if cultured without secondary stimulation.
Genetic Control of Lymphocytes Suppression
Authors: Primi Daniele , Hammarström Lennart , Smith C. I. Edvard
Published: December 2022

Con A-activated cells from old NZB mice were found capable of inhibiting the polyclonal response of cells from young NZB and BALB/c animals. Furthermore, Con A-preactivated spleen cells from young NZB and BALB/c mice did not significantly affect the response of spleen cells from old NZB mice. These results suggest that the defective suppressive activity in old NZB mice may be traced to a defect at the B cell level.
Functional Characteristics of BALB/c T Cell Lines: Suppression in the Mixed Lymphocyte Response and Cell-Mediated Lysis
Authors: Kim K. Jin , Weinbaum Fredric I. , Asofsky Richard
Published: December 2022

Previously, 10 BALB/c T cell lines (Thy 1.2+, Ig-) were shown to express different combinations of Ly 1 and Ly 2 antigens. The possible immunologic function(s) of these tumor cells was determined by investigating the effects of these cells on the responses to mitogens, the mixed lymphocyte response (MLR), and the generation of cell-mediated lysis (CML) by normal spleen cells. Five T cell lines, Pl798 and BALENTL 3, 5, 8, and 9, continued to synthesize DNA after exposure to large doses of irradiation. Only BALENTL 4, 6, 7, and 14 (Ly 1-2+) and BALENTL 13 (Ly 1+2-) were radiosensitive and therefore amenable to study. BALENTL 4 and 14 gave significant suppression of the MLR between BALB/c and C57BL/6; BALENTL 14 also inhibited the generation of BALB/c effector cells against C57BL/6 spleen cells. None of these T cell lines had any effect on the proliferative response of BALB/c spleen cells induced by concanavalin A. However, there was approximately 50% suppression of the phytohemagglutinin response of BALB/c spleen cells by BALENTL 14.
Modulation of Immune Response by Bacterial Lipopolysaccharide (LPS): Multifocal Effects of LPS-Induced Suppression of the Primary Antibody Response to a T-Dependent Antigen
Authors: Uchiyama Takehiko , Jacobs Diane M.
Published: December 2022

Spleen cells from mice injected with 2 to 50 µg bacterial lipopolysaccharide (LPS) have a reduced capacity to make an antibody response in vitro to trinitrophenylated sheep erythrocytes (TNP-SRBC) when tested 1 to 7 days later. Recovery is gradual, and these cells are fully functional 2 weeks after in vivo LPS treatment. Unresponsiveness resides in the nonadherent splenic cell populations, and can be shown to have a suppressive cell component, which is irradiation sensitive and has some characteristics of a thymus-derived lymphocyte (T cell). In addition, neither bone marrow-derived lymphocytes (B cells) nor T cells in the spleens of LPS-treated mice are functionally normal in their abilities to cooperate during an antibody response in vitro. LPS-B cells cooperated poorly with nylon wool-enriched T cells from normal mice but cooperated well with irradiated carrier-primed T cells or nylon wool-purified splenic T cells from carrier-primed mice. LPS-T cells have a reduced capacity to interact with normal B cells and appear to contain a suppressor cell component. These results indicate that the effects of exposure of immunocompetent cells to LPS are multifocal and can include suppression as well as stimulation of antibody formation.
Distribution and Identity of Thy-1-Bearing Cells during Ontogeny in Rat Hemopoietic and Lymphoid Tissues
Authors: Ritter Mary A. , Gordon Lance K. , Goldschneider Irving
Published: December 2022

The distribution and identity of Thy-1+ cells during ontogeny in the rat were examined by immunofluorescent and histologic techniques, and the relative amount of Thy-1 antigen on different cell types was assessed by quantitative absorption. Thy-1+ cells were subsequently studied by dual immunofluorescence for the simultaneous expression of either T or B lymphocyte markers. The results indicate that the Thy-1 antigen is present on hemopoietic precursors and on at least some of their lymphoid (both T and B) and nonlymphoid descendants. In addition, six serologically distinct subpopulations of lymphoid cells were identified, and their position in T and B cell maturational pathways is discussed.
Immunoregulation of Heymann's Nephritis
Authors: Litwin Allen , Bash Jerry A. , Adams Louis E. , Donovan Rebecca J. , Hess Evelyn V.
Published: December 2022

Pretreatment of Lewis rats with a series of injections of a renal tubular antigen (RTA) in IFA prevented induction of Heymann's nephritis (HN) when the rats were challenged with RTA in FCA. This absence of disease was confirmed by immunofluorescent staining for rat IgG and histologic examination of the kidneys as well as by lack of development of significant proteinuria. Passive transfer of spleen and lymph node cells from rats receiving such pretreatment into syngeneic recipients prevented induction of HN when these recipients were challenged with RTA in FCA. Passive transfer of serum obtained from pretreated rats was without effect. These results suggest that one of the mechanisms involved in preventing HN by this pretreatment regimen was the induction of suppressor cells. The results of spleen cell transformation indicated that the suppressor cells were specific for RTA as the immune response to a second antigen, PPD, was unaffected. When rats already had active early HN, the disease course was unaffected by transfer of suppressor cells.
Selective Cell Surface Expression of Thymus Leukemia Antigen during S Phase of the Cell Cycle
Authors: Williams Jackie L. , Pickering Jerry W. , Wolcott Michael
Published: December 2022

The expression of the thymus leukemia antigen (TL) was studied on a murine leukemia cell line (ASL-1W) grown in vitro and separated into cell cycle phases by velocity sedimentation or growing synchronously in culture. The expression of TL was determined qualitatively by direct cytotoxicity and quantitatively by a modified inhibition of cytotoxicity assay. TL expression was found to vary with DNA synthesis. The hypothesis that expression is coordinately regulated with DNA synthesis, and the relationship of this to the restricted expression of TL on rapidly dividing cells is discussed.
Action of the C3b-Inactivator on Cell-Bound C3b
Authors: Law S. K. , Fearon Douglas T. , Levine R. P.
Published: December 2022

The action of C3bINA and β1H on cell-bound C3b is described in this paper. The α-polypeptide of C3b that binds covalently to cell surfaces is cleaved by the C3bINA and β1H into two fragments: one of 60,000 (C3bα-60) and another of 40,000 (C3bα-40) daltons. The β-chain of C3b is unaffected by the C3bINA and β1H. The three polypeptides, C3bα-60, C3bα-40, and C3β, are held together as a single unit by disulfide bonds. This unit, referred to as C3b', is covalently bound to cell surfaces via the C3bα-60 polypeptide. The conversion of C3b to C3b' by C3bINA and β1H abolishes the ability of the C3b-bearing cells to adhere to human erythrocytes as well as the ability to form, on the cell surface, the B, &D, and properdin-dependent amplification C3-convertase. However, the agglutinability of the cells with either anti-C3c or anti-C3d is not affected.

Treatment of the C3b'-bearing cells with trypsin releases fragments of C3b' into solution, leaving a polypeptide of 32,000 daltons covalently linked to the membrane. Since the trypsinized cells are agglutinable by anti-C3d but not by anti-C3c, the 32,000 dalton polypeptide appears to correspond antigenically to C3d.
Activation of Membrane Phospholipase a by Guinea Pig Lymphotoxin (GLT)
Authors: Kobayashi Yoshiro , Sawada Jun-Ichi , Osawa Toshiaki
Published: December 2022

The effect of GLT on target cells (L·P3 cells) labeled with radioactive fatty acids was examined. After incubation for 1 or 2 hr, no change in the survival ratio was observed, but radioactive fatty acid was released from phospholipids of the cells. When incubation was continued for 4 or 5 hr, the survival ratio was decreased with a concomitant increase of the radioactivity of triglyceride of the target cells and enhancement of calcium uptake into the target cells. These results could be explained by activation of membrane-bound phospholipase A after the binding of GLT to cell surface receptors.
Suppression of Cell-Mediated Immune Reactions by Monosaccharides
Authors: Baba Tohru , Yoshida Tomoko , Yoshida Takeshi , Cohen Stanley
Published: December 2022

Various monosaccharides have been shown capable of inhibiting lymphokine activity in in vitro assay systems. In this study, we demonstrate that l-fucose can inhibit the ability of lymphokine-containing supernatants to induce skin reactions or cause reductions in the macrophage content of peritoneal exudates. Moreover, l-fucose can inhibit the cutaneous delayed hypersensitivity reaction and the peritoneal macrophage disappearance reaction (MDR) induced by antigen in actively immunized guinea pigs. The effect of l-rhamnose, another sugar with in vitro inhibitory activity, was investigated in the MDR, and was also shown to be inhibitory. l-arabinose, which has no in vitro effect on lymphokines, had no suppressive effect on any of the in vivo systems studied. No monosaccharide inhibition of Arthus reactions or nonspecific inflammation could be found in these studies. The results demonstrate that monosaccharides capable of inhibiting lymphokine activity in vitro are effective in suppressing in vivo manifestations of cellular immunity.
IgA Anti-Dextran B1355 Responses
Authors: Kagnoff Martin F.
Published: December 2022

Injection of mice bearing the Ig-1a allotype with dextran Bl355 results in an IgM antibody response that is generally regarded as thymus independent. Moreover, the antibody is directed to α[1,3] determinants on dextran Bl355 and shares cross-reacting idiotypic determinants with a λ1 IgA (J558) myeloma protein as well as a λ1 IgM (MOPC 104E) myeloma protein. In this study, we show that BALB/c (Ig-1a) mice injected with dextran Bl355 produced highly significant IgA anti-dextran responses with specificity directed to the α[1,3] epitope. Kinetics of the IgA anti-dextran response in BALB/c mice paralleled kinetics of the IgM response. However, the magnitude of the IgA response was markedly T cell dependent and age dependent.
A Quantitative Assay for Experimental Allergic Encephalomyelitis in the Rat Based on Permeability of Spinal Cords to 125I-Human γ-Globulin
Authors: Stohl William , Kaplan Marc S. , Gonatas Nicholas K.
Published: December 2022

We have developed a quantitative assay for experimental allergic encephalomyelitis (EAE) in the rat based on permeability of the spinal cord to 125I-human γ-globulin (HGG). This assay is highly reproducible and eliminates many of the drawbacks of assaying for EAE on the basis of clinical and/or histologic criteria. Using the assay, we have shown a direct correlation between onset of histologic changes in the spinal cord and onset of permeability changes in the spinal cord. No rat without histologic lesions manifest permeability alterations, and all rats with histologic lesions did manifest increased permeability to 125I-HGG. Furthermore, strains of rats susceptible to EAE demonstrated permeability changes, whereas resistant rats did not. In addition, we demonstrated by permeability and histologic criteria that guinea pig myelin basic protein emulsified with incomplete Freund's adjuvant is encephalitogenic in the Lewis rat. We also demonstrated that recipients of passive transfer of sensitized cells develop permeability changes along with histologic lesions. We conclude that measuring permeability to 125I-HGG in the spinal cords of rats is a valid assay for EAE, and it improves upon current indices of EAE in that it is readily quantifiable.
Functional Activation of Immune Lymphocytes by Antigenic Stimulation in Cell-Mediated Immunity
Authors: Yamamoto Yusei , Onoue Kaoru
Published: December 2022

Histocompatibility-linked restriction of macrophage-T lymphocyte interaction in antigen-induced MIF production by sensitized lymphocytes was examined, by using combinations of inbred strain 2, strain 13, and JY-1 guinea pigs. The effective interaction of the antigen-bearing macrophages with the immune T lymphocytes was observed when the donor of the antigen-bearing macrophages and that of the immune lymphocytes shared Ia antigens of the major histocompatibility complex. Identities of B antigens and S antigens were not important for this cooperation. It was further demonstrated that the previously reported soluble factor derived from LPS-stimulated peritoneal adherent cells (macrophages) could help antigenic activation of the immune lymphocytes across the strain barrier provided a small number of macrophages (0.01%) from syngeneic strain were present. These results show that the presence of macrophages is absolutely required to present antigen to immune T lymphocytes in a genetically restricted manner and the soluble factor from macrophages appears to give a nonspecific effect on the lymphocyte activation in addition to or in collaboration with antigenic stimulation.
Evaluation of the Quantity and Affinity of Human IgG “Blocking” Antibodies
Authors: Adkinson N. Franklin , Sobotka Anne K. , Lichtenstein Lawrence M.
Published: December 2022

We have analyzed the quantity and avidity of IgG antibodies in human sera specific for allergenic proteins of clinical relevance. The methods used were saturation analysis, which provides quantitative estimates of antibody content and Scatchard and Sipps analyses of antigen-binding experiments, which provide estimates of antibody affinity and homogeneity and, by mathematical extrapolation, antibody quantity as well. We studied human IgG “blocking” antibody specific for three unrelated allergens: honeybee phospholipase A (PLA), ragweed antigen Ra5 (Ra5), and the benzylpenicilloyl (BPO) hapten.

Antibody content determined by saturation analysis was highly correlated (r = 0.99; p < 0.001) with estimates obtained by mathematical extrapolation of Scatchard and Sipps plots. The average equilibrium constant (K0) for IgG anti-PLA was 3 × 1010 M-1; for IgG anti-Ra5, the average K0 was 0.34 × 1010 M-1; and the K0 of a single serum with IgG anti-BPO had a high binding constant (12 ± 1.3 × 1010 M-1). The heterogeneity coefficients obtained from the Sipps analysis were consistently high for all allergens studied, suggesting remarkable homogeneity of antibody. Since antibody affinities varied over a relatively narrow range, it was possible to obtain estimates of antibody content by interpolation from the dilution curve of a standardized serum that agreed very well (r = 0.99; p < 0.001) with values obtained by the more laborious saturation analysis technique.

It appears that saturation analysis and the Scatchard and Sipps transformations can be usefully applied to the study of human IgG “blocking” antibodies against large and small m.w. allergens, as well as haptens. The average affinity constant of human IgG blocking antibody is high when compared with hyperimmune animal antisera. The affinity constant for each allergen system varied over less than a 10-fold range in the limited number of sera studied, suggesting that variability in K0 is not a major contributor to the allergen-binding capacity of most sera.
β2-Microglobulin Enhances Human Lymphocyte Surface Receptor Expression for IgG
Authors: Birch Ruth E. , Fanger Michael W. , Bernier George M.
Published: December 2022

Because of its close sequence homology to immunoglobulin constant domains, the effect of β2-microglobulin (β2µ) on the expression of Fc receptors on human peripheral blood lymphocytes was studied. Treatment of peripheral blood lymphocytes (PBL) at 4°C with β2-microglobulin resulted in an increase in the percentage of lymphocytes expressing receptors for the Fc portion of IgG (Fcγ) from 34% in untreated preparations to 54% in preparations incubated with 50 µg of β2µ/2 × 106 cells. As shown by the binding of 125I-labeled β2µ, the molecule was taken up by peripheral lymphocytes and was associated with those lymphocytes that expressed Fcγ receptors. β2µ treatment had no demonstrable effect on the expression of other lymphocyte markers—E, EAC, sIg. The increase in Fcγ receptor expression was observed on T cells. Removal of adherent cells had no effect on β2µ induced Fcγ receptor expression, but the increased Fcγ receptor expression was abrogated by treatment with rabbit anti-human β2µ F(ab′)2 fragments. These findings suggest that high concentrations of β2µ exert effects on lymphocyte surfaces that may have functional as well as structural consequences in regulation of the immune system.
Molecular and Quantitative Analysis of Helper T Cell-Replacing Factors on the Induction of Antigen-Sensitive B and T Lymphocytes
Authors: Watson James , Aarden Lucien A. , Shaw Jennifer , Paetkau Vermer
Published: December 2022

Murine spleen cells activated by the T cell mitogen concanavalin A (Con A) secrete factors that exhibit biologic activity on both bone-marrow (B) and thymus-derived (T) lymphocytes. In the presence of heterologous erythrocyte antigens, these factors stimulate antibody synthesis in T cell-depleted spleen cultures. In the presence of Con A, these factors stimulate mitogenic responses in thymocyte cultures where the cell density is too low to support mitogenic responses to Con A alone. Both culture systems have been developed as quantitative assays for the activities of the Con A factors. Purification of the biologic activities by salt precipitation, gel filtration, ion-exchange chromatography, and isoelectric focusing (IEF) establish that the molecules responsible for effects on B and T lymphocytes in these assay systems are identical. The quantitative assays performed reveal that these factors are active in each culture system at concentrations below 10-9 M. The IEF-purified factors also stimulate the specific generation of cytotoxic lymphocytes to allogeneic tumor cells in culture. These experiments reveal a requirement for helper T cells in the induction of antibody synthesis in B lymphocytes, mitogenic responses to Con A in T lymphocytes, and cytotoxic T cell precursors to effector cells. Each assay system may be a measure of the induction of antigen-sensitive lymphocytes, suggesting that B and T lymphocytes respond to similar, if not identical, helper T cell activity.
Complement Lysis of Resealed Red Cell Membrane Ghosts Pretreated with Glutaraldehyde
Authors: Giavedoni Elda B. , Chow Yeh Mei , Dalmasso Agustin P.
Published: December 2022

To investigate the effect of extensive cross-linkage of membrane proteins on the susceptibility of the membrane to complement (C) lysis, resealed sheep erythrocyte membrane ghosts (M) containing 3H-inulin or 3H-Dextran 20 were treated with glutaraldehyde (gl). The release of these marker molecules by the action of antibody (A) and C was then measured and compared to controls not treated with gl. There was an increase in C-induced release over a wide range of gl concentrations with guinea pig or human serum but not with rabbit serum. Most experiments were carried out with 0.25% gl, which caused a profound increment in the rate of marker release. MglAC1-7 treated with human C8 and C9 underwent more release than MAC1-7, but the difference diminished with high C8 concentrations. Gl treatment did not alter the number of 125I-C8 molecules that bind to M containing the first seven C components. Upon Sephadex G-200 chromatography, the Dextran 20 released by C from MglA was found to contain large molecules that were not present in the material released from MA. Because gl causes extensive cross-linkage of the membrane proteins, we conclude that they play no essential role in the production of the primary C lesion. Therefore, in biologic membranes the lipid bilayer constitutes the primary site of the C attack mechanism, as shown previously with artificial membranes. The studies also demonstrate that gl-induced cross-linkage of proteins and certain phospholipids results in membranes that upon reaction with C develop larger or more stable primary C lesions than native membranes.
Characterization of the Spontaneous Murine B Cell Leukemia (BCL1)
Authors: Vitetta E. S. , Yuan D. , Krolick K. , Isakson P. , Knapp M. , Slavin S. , Strober S.
Published: December 2022

We have raised an anti-idiotypic antibody against the cell surface IgM of the murine BCL1 tumor cells. This antiserum reacts exclusively with the IgM expressed on the tumor cells and detects a unique population of cells in the spleen and blood of the tumor-bearing mice. When these cells are stimulated in vitro with LPS, they secrete an IgM bearing the same idiotype as the cell surface Ig. These results are discussed in terms of a model for the immunotherapy of a chronic lymphocytic leukemia-like syndrome in mice.
Selective Adherence of Lymphocytes to Myelinated Areas of Rat Brain
Authors: Kuttner Barry J. , Woodruff Judith J.
Published: December 2022

An in vitro system developed for studying lymphocyte binding to high endothelial venules (HEV) of lymph nodes was used to determine if there are similar binding sites in other organs of the rat. Thoracic duct lymphocytes (TDL) adhered selectively and uniformly to white matter when overlaid onto glutaraldehyde-fixed tissue sections of cerebellum and cerebrum. The pattern of TDL adherence to cerebellar sections showed that binding to nonmyelinated areas was negligible. Comparison of TDL-white matter to TDL-HEV binding demonstrated that the density of adherence to each site was quantitatively similar. In contrast, lymphocytes exhibited little tendency to bind to tissue sections of liver, spleen, heart, thymus, and salivary glands. TDL adherence to cerebellar white matter occurred rapidly, was cell dose dependent and optimal at 7°C. White matter binding was also a property of spleen lymphocytes but the thymus was deficient in cells with this capability. The affinity of TDL and spleen lymphocytes for myelinated areas of the brain suggests the presence of myelin binding receptors on these cells.
Carrier-Specific Immune Memory to a Thymus-Independent Antigen in Congenitally Athymic Mice
Authors: Schott Chester F. , Merchant Bruce
Published: December 2022

Immune memory to the DNP epitope coupled to a nonmitogenic thymus-independent carrier, Ficoll, was demonstrated in congenitally athymic outbred Swiss mice. Strong IgM and modest IgG components of this memory were detected. Moreover, this memory was carrier specific since it was elicitable only when DNP-Ficoll primed mice were challenged with DNP-Ficoll and not when similarly primed mice were challenged with DNP coupled to pneumococcal polysaccharide or to keyhole limpet hemocyanin. Ficoll primed mice also demonstrated a memory response when challenged with DNP-Ficoll. These findings indicate that a non-T cell, presumably a B cell, is capable of recognizing the carrier epitopes of this thymus-independent hapten-carrier complex.

Unlike their athymic counterparts, euthymic mice of the same genetic background failed to demonstrate the IgM component of this memory, but they did demonstrate modest carrier-specific IgG memory. These results strongly suggest that suppressor T cells are either directly or indirectly important in regulating the IgM memory response of these mice to DNP-Ficoll. Indirect regulation could possibly occur via an antibody-mediated specific immune suppression mechanism.