bluuebunny/crossref_metadata_2025 huggingface dataset:
Natural Cytotoxic Reactivity of Human Lymphocytes Against a Myeloid Cell Line: Characterization of Effector Cells
Authors: West William H. , Cannon Grace B. , Kay H. David , Bonnard Guy D. , Herberman Ronald B.
Published: December 2022

Using a series of techniques to identify and deplete various peripheral blood lymphocyte subpopulations, we studied the cytotoxic reactivity of normal individuals against the myeloid cell line K-562 in a 4-hr 51chromium-release assay. Depletion of lymphocytes bearing complement receptors had a variable, usually negligible effect on cytotoxicity. In contrast, depletion of lymphocytes bearing Fc receptors abrogated target cell lysis. Separation of lymphocytes with high-affinity binding of sheep red blood cells (SRBC) evidenced by rosette formation at 29°C yielded a population of rosette-forming cells containing few cytotoxic cells, whereas separation of total E-RFC under optimal rosetting conditions produced a rosette fraction containing a major proportion of the effector cells. These data indicate that the cytotoxic lymphocyte in this system is Fc receptor positive, largely complement receptor negative, and may possess low density or low affinity receptors for SRBC.
Induction of Granulomatous and Elicitation of Cutaneous Sensitivity by Partially Purified SEA of Schistosoma mansoni
Authors: Boros Dov L. , Tomford R. , Warren K. S.
Published: December 2022

Schistosomiasis mansoni is a helminth infection of about 80 million people in the tropics. Granulomas which appear around schistosome eggs disseminated in the tissues of the definitive host are primarily responsible for the occurrence of hepatosplenic disease (1). This granulomatous reaction has been shown to be a specific, cell-mediated host response (2) to soluble antigens secreted by the living, mature schistosome eggs (3). Furthermore, when soluble egg antigens (SEA)2 obtained by homogenization and ultracentrifugation were administered in microgram amounts without adjuvant they induced delayed hypersensitivity in mice and guinea pigs (3, 4). When SEA was coated onto bentonite particles it elicited typical granulomas in specifically sensitized animals (5, 6). Crude SEA was also used in delayed skin tests (3, 4) and in the stimulation of blastogenesis (7) and lymphokine production (8, 9) in cultures of sensitized lymphocytes.
Structural Analysis of the Peptides Derived from Specific Acid-Catalyzed Hydrolysis at Aspartylprolyl Peptide Bonds in Human J Chain
Authors: Mole John E. , Bhown Ajit S. , Bennett J. Claude
Published: December 2022

Human J chain from IgM has been selectively cleaved at three aspartylprolyl peptide bonds to yield four fragments containing 62, 20, 25, and 22 amino acids, respectively. The amino acid sequence of each peptide has been partially determined, (59 of a total of 129 residues) and its position in the J chain ascertained. There were no obvious similarities to known sequences in other immunoglobulin polypeptide chains.
Bidirectional Amplification of Macrophage-Lymphocyte Interactions: Enhanced Lymphocyte Activation Factor Production by Activated Adherent Mouse Peritoneal Cells
Authors: Meltzer Monte S. , Oppenheim Joost J.
Published: December 2022

Adherent peritoneal cells (90 to 95% macrophages) from mice injected with Mycobacterium bovis, strain BCG, or certain noninfectious agents such as pyran copolymer or phytohemagglutinin showed increased chemotactic and tumoricidal capacity in vitro. These activated macrophages elaborated 2 to 5 times more lymphocyte-activating factor (LAF) in vitro than equal numbers of adherent cells from untreated mice. In contrast, adherent PC from mice treated with thioglycollate or mineral oil were not cytotoxic and did not produce more LAF than PC from untreated mice. Adherent PC from untreated nude mice, which have increased chemotactic and tumoricidal capacity in vitro, also exhibited enhanced LAF production compared to adherent PC from their normal littermates. Increased production of LAF was also evident with adherent PC and the macrophage-like tumor cell line P388D1 after incubation in vitro with bacterial endotoxins or with antigen-induced lymphokines. These data indicate that adherent PC can be activated either in vivo or in vitro to elaborate more LAF. Thus, activated macrophages are more effective than normal macrophages in amplification of the afferent limb of immune responses as well as in their effector functions.
Isolation of the Third Component of Mouse Complement
Authors: Gyöngyössy Maria I. C. , Assimeh Seth N.
Published: December 2022

The third component of mouse complement has been purified to homogeneity from pseudoglobulin by two chromatographic steps involving sequential cation- and anion-exchange chromatography. Mouse C3 is a β-glycoprotein of m.w. 210,000 consisting of two nonidentical, disulfidelinked polypeptide chains of m.w. 130,000 and 75,000, respectively. The properties of this molecule and its proteolytic fragments closely resemble those of human C3.
Quantitative Studies on the Precursors of Cytotoxic Lymphocytes
Authors: Teh H.-S. , Phillips R. A. , Miller R. G.
Published: December 2022

A microculture system was used to examine the specificity of cytotoxic T lymphocytes (CL) derived from single precursors. Our data indicate that clones derived from single precursors of CL (CLP) are specific. Thus, RNC mice (H-2k) simultaneously stimulated by both H-2d and H-2b antigens produce clones which are specific for either H-2b or H-2d but not for both. In lymph node (LN), the frequency of CLP giving rise to clones specific for H-2d and H-2b is 1/480 and 1/860, respectively. Clonal specificity is also maintained when Con A is used to activate the precursors. With a nonspecific assay of CL, it was found that 2.5% of the cells in LN are stimulated by Con A to produce clones of CL. Among the CLP stimulated by Con A, 2.1% of the clones are specific for H-2d alloantigens. An unexpected finding was that 18% of the clones produced by Con A stimulation lysed EL4 tumor cells, which carry H-2b; however, only a small proportion of these clones were specific for the H-2b alloantigen. The antigen on EL4 which accounts for the high response has not been characterized.
Antigen-Initiated B Lymphocyte Differentiation
Authors: Fidler John M. , Howard Maureen , Schlegel Robert A. , Vadas Mathew , Shortman Ken
Published: December 2022

The characteristics of memory B cell antibody-forming cell (AFC) progenitors from long-term hapten-primed CBA mice were investigated by using sedimentation velocity and buoyant density separation to isolate physically distinct B cell sub-sets. The isolated fractions were assayed by the adoptive immune response to NIP-POL antigen, under conditions where neither T cells nor other accessory cells were limiting the IgM or IgG AFC responses. The results were compared to previous studies on the IgM AFC-progenitors of unprimed adult mice.

Splenic IgM and IgG memory AFC-progenitor activity was largely found among the typical B cells of slow to medium sedimentation rate, in contrast to the faster sedimenting IgM AFC-progenitor activity of unprimed animals. Splenic IgM and IgG memory AFC-progenitor activity was found among the medium to light density cells, and so resembled by this parameter the IgM AFC-progenitor activity in unprimed animals. Thoracic duct lymphocytes from hapten-primed mice also exhibited memory IgM and IgG AFC-progenitor activity in the slow-medium sedimentation range. However, in contrast to spleen, the IgM and IgG memory AFC-progenitor activity in lymph was found among very dense B cells.

Two physically distinct sub-populations of memory B cells have thus been identified, namely: i) small, medium-light density, presumably tissue-resident B lymphocytes found in spleen; and ii) small, dense, presumably recirculating B lymphocytes found in lymph. Both physical forms include IgM and IgG progenitors. Both forms are distinct from the larger, medium-light density “virgin” AFC-progenitors in the spleen of unprimed adult mice.
Differentiation of B Lymphocytes in C3H/HeJ Mice: The Induction of Ia Antigens by Lipopolysaccharide
Authors: Watson James
Published: December 2022

The lipid A moiety of bacterial lipopolysaccharide (LPS) elicits several types of responses in murine B lymphocytes. First, lipid A induces the nonproliferative expression of cell surface antigens in more immature cell types. Second, lipid A induces a mitogenic response in more mature B cell types. Lipid A induces the expression of Ia antigens on bone marrow cells from C3H/DiSn but not C3H/HeJ mice. The Ia-inducible cells possess surface immunoglobulin. Agents that elevate intracellular levels of adenosine 3',5'-cyclic monophosphate (cyclic AMP) induce the appearance of Ia antigens on B lymphocytes from both C3H/HeJ and C3H/DiSn mice, suggesting that lipid A exerts its inductive effects by increasing cyclic AMP levels in cells. In contrast to what is observed by using other strains of mice, mature B lymphocytes from C3H/HeJ mice do not support a mitogenic response to lipid A. The subpopulation of B lymphocytes in C3H/HeJ mice that normally respond mitogenically to LPS not only appear to lack an LPS-response mechanism utilized in the mitogenic pathway, but they lack the LPS-response pathway of the immature B cell types. A lipid A-bound protein (LAP) induces both the expression of Ia and a mitogenic response in the different subpopulations of B lymphocytes from C3H/HeJ and C3H/DiSn mice. The genetic defect in C3H/HeJ mice that limits responses to lipid A may be associated with a receptor that is normally expressed on many different cell types.
Suppression of Adult Rat Lymphoid Proliferative Responses by Homologous Neonatal Serum
Authors: Weiss Arthur
Published: December 2022

The ontogeny of the immune response has usually been studied in intact neonatal animals or in cells isolated from neonatal animals (1–4). These studies have demonstrated an age-dependent emergence of immune responsiveness. Lymphoid cells isolated from rats soon after birth have a reduced capacity to respond to mitogens or to alloantigens in mixed leukocyte culture (MLC)3 (5, 6). Mechanisms controlling the development of immunologic responsiveness in lymphoid cell populations are poorly understood.

Extracellular materials may influence the reactivity of lymphoid cells. For example, factors present in normal serum are potent inhibitors of MLC (7, 8). Many of these serum factors have been isolated from the α-globulin fraction (9); one of these, purified α-fetoprotein, which is the first major serum protein to appear in the embryo, has exhibited immunosuppressive capacity in MLC and mitogen studies (10–12).
A Simple One-Step Hemolytic Assay for C2 with C2-Deficient Human Serum
Authors: Ngan Bo-Yee , Gelfand Erwin W. , Minta Joe O.
Published: December 2022

A simple one-step procedure has been developed for the molecular titration of C2 by utilizing the ability of the test material to restore the hemolytic activity of human serum selectively deficient in C2 (C2D serum). In this assay, equal volumes of EA (108 cells/ml), C2D serum (1/20), and a suitable dilution of a source of C2 were incubated at 37°C for 60 min and the fraction of cells lysed was used to calculate the effective molecules of C2/ml test material. The assay can be used to titrate C2 in human, guinea pig, rat, mouse and rabbit sera, but not C2 in dog serum. The assay is simple and reproducible and comparable in sensitivity to the conventional two-step assay with EAC14 cells and Cgp-EDTA.
Histamine-Induced Inhibition of Normal Human Basophil Chemotaxis to C5a
Authors: Lett-Brown Michael A. , Leonard Edward J.
Published: December 2022

Histamine in concentrations as low as 10-8 M inhibited the chemotactic response of normal human basophils to C5a. Histamine had no effect on basophil chemotaxis to lymphocyte-derived chemotactic factor. Histamine inhibition of basophil chemotaxis to C5a was prevented by metiamide, a drug which blocks H2 receptors for histamine. Since the accumulation of basophils in delayed cutaneous basophilic hypersensitivity (CBH) reactions may occur in part because of chemotaxis to C5a, and since C5a can induce histamine release, histamine inhibition of chemotaxis may limit basophil infiltration in CBH lesions.
Human Immunodeficiency Disease: Impairment of Cellular Interactions Leading to Abnormal Mediator Production in Mixed Lymphocyte Culture Reaction
Authors: Gorski Andrzej J. , Dupont Bo , Hansen John A. , Safai Bijan , Pahwa Savita , Good Robert A.
Published: December 2022

Leukocyte migration inhibitory factor (LMIF) production in mixed lymphocyte culture (MLC) reactions is the result of cellular interactions based on two separate phenomena: the capacity of lymphocytes to stimulate in MLC, and the capacity of lymphocytes to respond in MLC. Puromycin-treated lymphocytes are capable of stimulating allogeneic cells for LMIF production, but are unable to respond with synthesis of LMIF (one-way MLC-LMIF test). We have studied the stimulating and responding capacity of lymphocytes from patients with different immunodeficiency syndromes in a one-way MLC-LMIF assay. Lymphocytes from patients known to have qualitative and quantitative defects of T cell or B cell functions (Hodgkin's disease, mycosis fungoides, thymoma, chronic lymphatic leukemia) were found to respond poorly as measured by mediator production although their stimulating function was frequently retained. Patients with advanced solid tumors often had both MLC-stimulating and responding functions depressed. There was no apparent correlation between mitogen response and MLC-induced LMIF responses or between MLC proliferative response (as measured by thymidine incorporation) and mediator production. Studying of stimulatory and responding capacity of lymphocytes in the MLC-LMIF assay provides a new tool for assessing immunocompetence and allows for in vitro evaluation of cellular interactions that may play an important role in vivo.
Effects of Lymphocytes from Marek's Disease-Infected Chickens on Mitogen Responses of Syngeneic Normal Chicken Spleen Cells
Authors: Theis Gail A.
Published: December 2022

PHA responses have been measured in lymphoid cell cultures prepared by mixing normal chicken spleen cells with spleen or thymus cells from syngeneic chickens infected with the oncogenic herpesvirus MDV. Results of these studies may be summarized as follows: 1) spleen cells from MDV-infected chickens with visceral lymphomas inhibit the PHA response of normal spleen cells possibly by release of soluble inhibitory factors in response to the mitogen; 2) lymphoid cells from asymptomatic MDV-infected chickens, although hyporeactive themselves to PHA, can have a stimulatory effect on PHA responses of normal spleen cells in mixed cultures; 3) spleen cells from MDV-infected chickens, effectively protected from viral oncogenesis by HVT vaccination, show normal reactivity to PHA in separate cultures and may react in mixed cultures like normal lymphocytes, with neither a pronounced stimulatory nor inhibitory effect on the PHA response of normal spleen cells.
Cell-Mediated Immune Responses in Recurrent Herpesvirus Infections
Authors: Lopez Carlos , O'Reilly Richard J.
Published: December 2022

Studies in immunosuppressed and immunodeficient patients indicate that the cell-mediated immune response appears to be responsible for controlling reactivated herpesvirus infections. In this study, the various parameters of a herpesvirus (types 1 and 2) antigen specific lymphocyte proliferation assay were optimized and used to evaluate individuals with clinical, recurrent HSV-1 and HSV-2 infections. Normal individuals with neutralizing antibody to HSV-1 or HSV-2 responded to virus antigen in culture as well as individuals with recurrent disease. Normal individuals without neutralizing antibody responded with a significantly lower response. Specificity of the lymphocyte proliferation assay was observed most strikingly in normal individuals with a rare HSV-1 infection during the vesicular eruption. Specificity was also observed by determining the ratio of the response to HSV-1 as compared to the response to HSV-2. Evaluated in this manner, individuals with recurrent HSV-1 infections had significantly higher ratios than individuals with HSV-2 infections and vice versa. Data from individuals with recurrent disease was compared to that of normal individuals to determine whether the former demonstrated a specific alteration in this response. Individuals with recurrent disease were found to have higher neutralizing antibody titers than normals. The neutralizing antibody titers in normal individuals correlated well with the lymphocyte proliferation assay results, whereas a similar evaluation in individuals with recurrent disease gave a negative correlation. The ratio of HSV-1 response/HSV-2 response also demonstrated a suppressed response in recurrent infections to the homologous virus during active disease, which disappeared when the individual was convalescent. These studies indicate that individuals with recurrent HSV infections have virus antigen specific alterations of their cell-mediated immune response, which can be associated with their disease.
Resistance to Murine Leukemia in Mice Receiving Simultaneous Injections of Syngeneic Hybrid and Parental Neoplastic Cells
Authors: Liang Weitze , Cohen Edward P.
Published: December 2022

(A/J × C3H/HeJ)F1 mice (H-2a/k) exhibit no apparent resistance to the malignant proliferation of ASL-1 leukemia cells (H-2a), a spontaneously occurring neoplasm of A/J mouse origin. The period of survival after injections of ASL-1 cells is related to the number of cells introduced into the susceptible host. (A/J × C3H/HeJ)F1 mice receiving viable ASL-1 cells along with ASL-1 × LM(TK)- somatic hybrid cells (H-2a/k), syngeneic with the recipients, survived for statistically significant (p < 0.001), longer periods than did mice receiving ASL-1 cells alone. Mice receiving ASL-1 and ASL-1 × LM(TK)- hybrid cells together survived for approximately twice as long as animals receiving ASL-1 cells alone. Mice receiving ASL-1 cells followed by hybrid cells survived for longer periods as well, but the therapeutic effects were less successful. ASL-1 × LM(TK)- hybrid cells are rejected after forming localized tumors by the immunocompetent F1 mice. In some instances, F1 animals previously rejecting hybrid cells and challenged subsequently with ASL-1 cells along with a second injection of hybrid cells survived indefinitely. The passive transfer of partial immunity to the leukemic cells could be achieved by injections of spleen cells from F1 mice that had rejected hybrid cells previously. Similar results were obtained for (A/J × C3H/HeJ)F1 mice receiving RADA-1 cells (H-2a), a radiation-induced leukemic line of A/J origin, along with RADA-1 × LM(TK)- hybrid cells (H-2a/k). The partial immunity induced by these lines of hybrid cells was cross-reactive: mice injected with ASL-1 cells and RADA-1 × LM(TK)- hybrid cells survived for prolonged periods, and vice versa.
Interaction with Homologous Erythrocytes of Rat T Cells Which Act as Aggressors in the Mixed Lymphocyte Reaction
Authors: Owen Frances L. , Stux Stephen V. , Nisonoff Alfred
Published: December 2022

Substantial percentages of T-enriched spleen lymphocytes or thymocytes of inbred rats were found to form rosettes with the RBC of homologous strains. When an excess of RBC was used, essentially all of the rosette-forming subpopulation of lymphocytes was removed when the rosettes were separated by centrifugation. After depletion of the lymphocytes, reactive with RBC of one strain, most of the lymphocytes reactive with RBC of other strains could be recovered in the supernatant. A very large percentage of lymphocytes of the BN strain formed rosettes when a mixture of the RBC of five other strains was tested; the percentage was, however, somewhat lower than that predicted on the basis of complete additivity. Rosettes dissociated when warmed to 37°C. The lymphocytes recovered were unable to form rosettes again. In nearly all instances, the subpopulation that formed rosettes with RBC of a given strain included essentially all of the lymphocytes that acted as aggressors against peripheral leukocytes or mytomycin C-treated thymocytes of that strain; the lymphocytes in the supernatant always retained activity as aggressors against the leukocytes of one or more other strains and retained their responsiveness to PHA. Lymphocytes recovered from rosettes, by warming to 37°C, were highly reactive as aggressors in the MLR against the strain providing the RBC. Varying degrees of reactivity were noted against leukocytes of other strains.
Induction of Cell-Mediated Immunity to Chemically Modified Antigens in Guinea Pigs
Authors: Dailey Morris O. , Post Warren , Hunter Robert L.
Published: December 2022

Protein antigens covalently conjugated with lipid groups (dodecanoic acid) have previously been shown to stimulate strong delayed-type hypersensitivity (DTH) without the aid of adjuvants. The present experiments show that lipid-conjugated bovine serum albumin (L-BSA) is taken up in vitro by macrophages (Mø) 25- to 50-fold more than unconjugated BSA or aminidated BSA, neither of which induces DTH. Macrophages that take up 125I-labeled L-BSA in vitro stimulate DTH even more efficiently, when injected into syngeneic guinea pigs, than does soluble L-BSA. Tracer studies on the fate of radiolabeled BSA and L-BSA showed that much more L-BSA than BSA was retained by draining lymph nodes. Autoradiography demonstrated that 125I-L-BSA is rapidly taken up by Mø in the medullary sinuses of the lymph nodes. Some of this antigen is then transported into the paracortex, a region in which T lymphocytes predominate. The capacity of lipophilic antigens to stimulate cell-mediated immune responses may be caused by their increased uptake by Mø, resulting in more efficient presentation to immunocompetent T lymphocytes. The anatomical site of this Mø-T cell interaction may be within the sinusoids or paracortex of the draining lymph nodes.
Activity of Tumor-Associated Lymphoid Cells at Short Intervals after Administration of Irradiated Syngeneic and Allogeneic Tumor Cells
Authors: Schick Barbara , Berke Gideon
Published: December 2022

After a single intraperitoneal injection of irradiated tumor cells, host cells capable of responding against syngeneic tumors were detected in peritoneal exudates of mice. Although irradiation of the injected tumor prevented its overgrowth, it did not significantly alter the antigenicity of the tumor. Immunologic activities of tumor-associated host cells in the peritoneal cavity were continuously monitored, starting 48 hr after tumor administration. In vitro cell-mediated lysis of syngeneic tumors appeared as early as 3 days after irradiated tumor administration. In addition, peritoneal exudate cells from inoculated mice were capable of adoptively transferring immunity. Purification of these peritoneal exudate cells on nylon wool columns yielded a nonadherent Ig-negative lymphocyte fraction whose cytolysis was tumor-specific and T cell-associated. The macrophage-free lymphocyte fraction exhibited a higher in vitro activity against tumors than unpurified peritoneal exudates. This tumor-host system allowed the study of cells which directly interact with the tumor cells in vivo, starting shortly after tumor administration. The results reported in this paper show that tumor-associated lymphoid cells capable of mounting anti-tumor response in vivo and in vitro can be demonstrated as early as 3 days after tumor inoculation.
Inhibition of the Mixed Lymphocyte Culture Response with Anti-Lyb-4.1 Serum
Authors: Freund John G. , Ahmed Aftab , Dorf Martin E. , Sell Kenneth W. , Humphreys Robert E.
Published: December 2022

C57BL/Ks anti-L1210 serum, which recognized a non-H-2-linked B cell alloantigen, designated Lyb-4.1, specifically blocked the mixed lymphocyte culture (MLC) response to allogeneic cells that expressed the Lyb-4.1 determinant. Anti-Lyb-4.1 serum blocked the MLC response across H-2 and MLC disparities. To test that this effect was not the result of a toxic or nonspecific cell-coating action, the response of parental cells to F1 lymphocytes was studied in combinations in which only one parent expressed the recognized allele. MLC stimulation was blocked only when the responding parental cell recognized on the F1 cell H-2 or Mls disparities which were derived from the parent which possessed the Lyb-4.1 antigen.

Several DBA/2 tumors were characterized by cytotoxic and quantitation absorption assays for the presence of the B cell antigen. The presence of the antigen correlated with the ability of a limited number of tumors to stimulate the MLC response of H-2d identical BALB/c lymphocytes. An increased representation of the B cell alloantigen was found on the transformed B lymphoblast cell line in comparison to splenic B lymphocytes.
Generation of Cytotoxic T Lymphocytes during Coxsackievirus B-3 Infection
Authors: Wong Catherine Y. , Woodruff Judith J. , Woodruff Jack F.
Published: December 2022

The production of cytotoxic cells in the spleens of adult male BALB/c mice infected with Coxsackievirus B-3 has been examined. An in vitro 51Cr release assay was used to measure cytotoxic activity against virus-infected and uninfected neonatal syngeneic fibroblasts. Cytotoxicity of immune spleen cells against virus-infected targets was detected on the 3rd day after infection, reached a peak on day 7, and then declined to low levels by days 12 and 14. Spleen cells obtained 3 and 5 days after infection also exerted cytotoxicity against uninfected fibroblasts, but by the 7th day there was little or no reactivity against uninfected target cells, although activity against infected fibroblasts was maximal at this time. Reciprocal assays performed by using Coxsackie and vaccinia viruses provided evidence of virus specificity of the cytotoxic reaction. When spleen cells were obtained 7 days after infection, the Coxsackievirus-immune population was not cytotoxic for vaccinia-infected fibroblasts, and the vaccinia-immune population was not cytotoxic for Coxsackievirus-infected targets, although each immune cell preparation caused significant lysis of fibroblasts infected with the homologous virus. Additional studies showed that primary mouse or hyperimmune rabbit anti-Coxsackieviral serum could not block immune spleen cell cytotoxicity or induce complement-mediated lysis of in fected targets. The findings indicate that Coxsackievirus infection results in surface membrane alterations, but no evidence was obtained that antiviral antibody could react with the infected cells.
Topographic Determinants on Cytochrome c
Authors: Urbanski G. J. , Margoliash E.
Published: December 2022

Rabbit, mouse, and guanaco cytochromes c differ from each other by only two amino acid residues. The identification is described of all of the antigenic determinants of mouse and guanaco cytochrome c that elicit an antibody response in rabbits, and those of the rabbit and guanaco proteins that elicit antibodies in the mouse. All except one of these sites center around single amino acid residue differences between the antigen and the host cytochrome c. The corresponding antibody populations bind only to the areas of the protein in which the substitutions occur. Such antigenic determinants manifested in rabbits by guanaco and mouse cytochromes c are centered around residues 62 and 89, and residues 44 and 89, respectively. Similarly, the mouse recognizes sites containing residues 44 and 62 in guanaco cytochrome c, and residues 44 and 89 in rabbit cytochrome c. In none of these instances has a change in sequence failed to produce an antibody response. Each of these determinants appears to elicit and bind to its antibody, independently of other determinants present on the protein. In addition, two different autoantigenic responses have been detected. The antibodies produced against the determinant formed by glutamyl residue 62 of the guanaco protein in both rabbits and mice, the cytochromes c of which carry an aspartyl residue in that position, also bind to the aspartyl-containing region but with lower affinity. However, mouse and rabbit cytochrome c also elicit antibodies to the area of residue 62 in rabbits and mice, respectively, and these antibodies still bind more strongly to the glutamyl- than to the aspartyl-containing determinant. This last response occurs only when there are residue substitutions elsewhere in the molecule, because mice and rabbits fail to respond to their own cytochrome c. Antibodies produced in mice against the change from alanyl to valyl residue 44 by rabbit and guanaco cytochromes c also bind to the alanyl-containing determinant, except less tightly than to the valyl region. Conversely, antibodies raised in rabbits against the change from valyl to alanyl residue 44 only bind to this region when it carries an alanine. It is suggested that antigenic determinants that arise as a result of amino acid residue substitutions between the immunizing and the corresponding host protein, without a change in the spatial arrangement of the polypeptide backbone, be termed topographic determinants.
Radiosensitivity of T and B Lymphocytes
Authors: Anderson Robert E. , Olson George B. , Autry Johnna R. , Howarth John L. , Troup Gary M. , Bartels Peter H.
Published: December 2022

Groups of 10-week-old female CBA/J mice were exposed in whole body fashion to 0, 5, 50, and 500 rads and sacrificed in serial fashion 1, 3, 5, 7, 9, 15, and 30 days after irradiation for morphologic evaluation of thymus, spleen, lymph node, and Peyer's patch, and assessment of the relative numbers of thymus-derived (T) and bone marrow-derived (B) cells in these tissues. The absolute and relative numbers of recirculating T and B cells mobilizable by thoracic duct cannulation were also determined and compared with similar determinations with respect to peripheral blood lymphocytes.

B cell depletion occurred more quickly and was more pronounced in spleen and lymph node than T cell depletion at all three exposure doses. Depletion of T and B cells was roughly equal in peripheral blood and thoracic duct lymph. When present, regeneration of the T cell component occurred more rapidly than did B cell restoration. The latter often was incomplete at the time of the final sacrifice (day 30). PHA-responsive and Con A-responsive cells also appeared to differ with respect to the kinetics of cell death after whole body irradiation.
Neonatal Susceptibility to MHV3 Infection in Mice
Authors: Levy-Leblond E. , Dupuy J. M.
Published: December 2022

Up to 3 weeks of age, mice of the resistant A/J strain are fully susceptible to mouse hepatitis virus type 3 infection (MHV3). Immune deficiency, however, resulting from neonatal thymectomy or long term ALS administration led A/J animals to remain susceptible when tested at adult age. Whole spleen cells transferred from normal adult A/J donor mice protected suckling syngeneic recipients from i.p. infection with MHV3. Such a protective capacity of spleen cells was abolished after treatment with anti-θ serum and complement. Spleen cell separation by means of adherence to plastic also showed that neither the nonadherent nor the adherent populations injected separately were able to confer resistance to young mice challenged with the virus. Protection was not achieved with peritoneal cells originating from adult syngeneic animals. Transfer of resistance to MHV3 was obtained, however, when peritoneal cells were associated with adherent spleen cells.

This study indicated that two types of mature cells, at least, were required for transferring MHV3 resistance into newborn mice of the A/J strain: T lymphocytes and an adherent spleen cell population.
A Comparative Study of Detection Methods for Aleutian Disease Viral Antibody
Authors: Crawford Timothy B. , McGuire Travis C. , Porter David D. , Cho H. J.
Published: December 2022

Four methods of detecting and quantitating mink antibody against Aleutian disease (AD) virus were compared. Counterelectrophoresis, modified counterelectrophoresis, immunofluorescence, and complement fixation were performed blindly on 274 serum samples. All four methods were reliably specific for AD antibody. Immunofluorescence was less reproducible than the other systems. Immunofluorescence and complement fixation were 4- to 8-fold more sensitive than regular or modified counterelectrophoresis, but were limited by background staining and anti-complementary activity, respectively, when used to detect small amounts of antibody in undiluted sera.
A Microtiter Solid Phase Radioimmunoassay for Detection of the Human Reovirus-Like Agent in Stools
Authors: Kalica Anthony R. , Purcell Robert H. , Sereno Mitzi M. , Wyatt Richard G. , Kim Hyun W. , Chanock Robert M. , Kapikian Albert Z.
Published: December 2022

The development of a microtiter solid-phase radioimmunoassay for detection of the human reovirus-like (RVL) agent is described. The test is simple to perform and uses small amounts of reagents; a large number of specimens can rapidly be tested in a single test. Both filtered and unfiltered stool suspensions can be employed. The test was as sensitive as immune electron microscopy, and with an appropriate blocking test, was specific for the human RVL agent.
Modulation of Macrophage C3b Receptor Function by Cytochalasin-Sensitive Structures
Authors: Atkinson John P. , Michael J. Mark , Chaplin Hugh , Parker Charles W.
Published: December 2022

A quantitative rosette assay was employed in order to determine if through pharmacologic probes we could gain an insight into the nature of the interaction between C3b-coated particles and the macrophage C3b receptor. Rabbit alveolar macrophage monolayers were challenged with chromium-labeled, complement-coated (via cold agglutinin) human erythrocytes (HEC3b) and the per cent of bound counts determined in the distilled water lysate. With this assay system in which ingestion is negligible, the cytochalasins (A τ E τ D τ B) produced the most marked inhibition of rosette formation compared to control treated monolayers. No agent examined produced consistent augmentation. Cytochalasin A at 10-5, 10-6, and 10-7 M inhibited rosette formation by 77 ± 2, 44 ± 4 and 15 ± 7 (S.E.), per cent, respectively. Cytochalasin E was also markedly inhibitory. Cytochalasins B and D produced approximately 30% inhibition at 10-5 M. The cytochalasin effect was not secondary to an interaction between these agents and complement-sensitized erythrocytes, although cytochalasin E was also able to reduce erythrocyte-bound C3b reactivity. Cytochalasin A and E modulation of the macrophage C3b reactivity occurred within a few minutes and was only slightly reversible. Cytochalasins A and E could also disrupt preformed rosettes but the effect was not as pronounced as when these agents were present before and/or during the actual adherence phenomenon. Vinblastine and colchicine (10-5 and 10-6 M) also produced significant inhibition of rosette formation, although the magnitude of the effect was less than that for cytochalasins A and E. Further characterization of the vinblastine and colchicine effect demonstrated that the inhibition was rapid, irreversible over a 60-min incubation, and not explained by an alteration in macrophage attachment or in HEC3b reactivity. Agents producing insignificant inhibition of rosette formation included the following: dibutyryl cAMP and cAMP agonists (PGE1, theophylline), 8-bromo cGMP and cGMP agonists (carbachol, ascorbic acid), dimethylsulfoxide, heparin, ethanol, dextran sulfate, DEAE-dextran, and poly-L-lysine. The data suggest that cytochalasin, vinblastine and colchicine sensitive membrane structures, most likely microfilaments and microtubules, are important in the interaction of C3b-coated particles with the macrophage C3b receptor.
Membrane Transport by Murine Lymphocytes
Authors: Strauss Phyllis R. , Sheehan James M. , Kashket Eva R.
Published: December 2022

We have investigated membrane transport of thymidine and adenosine by bulk murine nonadherent spleen cells from animals treated with the lectin concanavalin A. Separation of cells from radioactive medium was achieved by means of the oil microfuge technique which permits incubation periods as short as 4 sec. We were able to distinguish between the membrane-linked function of transport and uptake which includes subsequent accumulation and metabolism of the transport solute.

Previously we reported that cells derived from untreated mice could not be shown to translocate thymidine in a carrier-mediated fashion. In contrast, cells from concanavalin A-treated mice showed two membrane transport systems for thymidine, a high affinity system (Km = 160 μM) and one with low affinity (Km = 4 mM). The transport of thymidine conformed to the usual criteria for membrane carrier-linked function: increased uptake with time, substrate saturability and chemical specificity. In contrast to thymidine uptake, the transport of adenosine by cells from lectin-treated mice was similar to that shown by cells from untreated animals. Thus a specific membrane-linked activity is altered differentially in the course of mitogen-effected lymphocyte stimulation. We have found that determination of DNA synthesis by the standard method gave values which were as much as 700% too high, since the counts obtained by direct precipitation with TCA of cells incubated with radiolabeled thymidine exceed the cell-associated radiolabel obtained by the rapid sampling technique. With lectin-stimulated cells, the discrepancy observed was up to three times greater. Hence the validity of the standard assay for blastogenesis must be viewed with caution.
Additional Rabbit IgA Allotypes, f69, f70, g76, g77: Control by the Cαf and Cαg Loci
Authors: Lammert John M. , Dray Sheldon , Knight Katherine L. , Hanly W. Carey
Published: December 2022

Genetic and immunochemical studies have led to the identification of four additional rabbit IgA allotypes controlled by the Cαf and Cαg loci. The following linkage combinations of the VHa and the “new” alleles were observed among the populations of rabbits studied: a1f70g76, a1f69g77, and a2f69g77. Cross-reactions among g74, g76, and g77 molecules with various anti-g anti-allotype antisera indicate that the IgA-g allotypic specificities are comprised of multiple antigenic determinants. These studies provide a basis for further understanding of the evolution and genetic control of the immunoglobulin heavy chain chromosomal region.
Genetic Control of the Humoral Response to an H-2 Public Specificity
Authors: Hansen Ted H. , Cullen Susan E. , Shinohara Nobukata , Schurko Elizabeth , Sachs David H.
Published: December 2022

The humoral response of mice to an H-2 public specificity, termed H-2. “28”, was found to be under genetic control. The genes determining this specificity were mapped to both the H-2K and H-2D regions, suggesting possible structural homologies between the products determined by these two regions. Genetic analyses indicated that a single non-H-2-linked gene regulates the anti-H-2. “28” response. In a backcross study, no linkage was detected between this putative H-2. “28” Ir gene and either the VH region or the Ly-2 locus to which the κ-light chain locus is linked. Thus, a regulatory rather than a structural genetic locus seems a more likely basis for differences in response to this antigen. Our data further indicate that control of the humoral response to H-2. “28” is determinant specific since responses of the same backcross mice to other K and D alloantigens were not found to be subject to the same control.
H-2 Homology Requirements for Secondary Cell-Mediated Lympholysis and Mixed Lymphocyte Reactions to TNP-Modified Syngeneic Lymphocytes
Authors: Schmitt-Verhulst Anne-Marie , Garbarino Carol A. , Shearer Gene M.
Published: December 2022

Secondary cell-mediated lympholysis (CML) and mixed lymphocyte reactions (MLR) were generated in a tissue culture system against trinitrophenyl (TNP)-modified murine syngeneic spleen cells. H-2 homology between primary and secondary TNP-modified stimulating cells was required in order to restimulate in the secondary CML. Strong proliferative responses (MLR) were detected only in the secondary cultures, for which H-2 homology was also required between TNP-modified primary and secondary immunogens. Intra-H-2 mapping for the secondary MLR indicated that the relevant regions of homology were I, D, and K and/or I-A. Homology throughout the entire major histocompatibility complex or at K plus I-A gave stronger MLR than did cultures in which there was homology between the primary and secondary phases at I or D only.
Regulation of Lipopolysaccharide-Induced Granulopoiesis and Macrophage Formation by Spleen Cells
Authors: Apte Ron N. , Hertogs Chanita F. , Pluznik Dov H.
Published: December 2022

Addition of bacterial lipopolysaccharide (LPS), a B cell mitogen, to mouse spleen cultures strongly stimulated production of colony-stimulating factor (CSF), the humoral regulator of granulopoiesis, and macrophage formation in vitro. Secretion of CSF from LPS-stimulated spleen cells coincided with cellular DNA synthesis and cell transformation and both activities could be attributed to the lipid A moiety of the molecule. Different experimental approaches were used to study the relationship of CSF release and lymphocyte activation in response to LPS: a) modification of LPS with polymyxin B, an antibiotic bactericidal for most Gram-negative bacteria, caused a marked reduction in mitogenic activity, although the ability to induce CSF was not significantly altered; b) spleen cells from CBA/N mice, a mutant strain with an x-linked genetic defect in immunologic and mitogenic responses to polyclonal activators including LPS, showed diminished mitogenic responses; however, high levels of CSF were produced; c) mitotic and DNA inhibitors (colchicine and cytosine arabinoside) did not affect CSF release although they completely inhibited mitogenicity. Thus, the spleen cell population participating in the process of LPS-induced CSF generation is probably a nondividing, terminally differentiated one without need for DNA synthesis. In addition, it was also shown that active RNA and protein synthesis are needed in this process.
Systemic Expression of Cutaneous Basophil Hypersensitivity
Authors: Dvorak Harold F. , Hammond M. Elizabeth , Colvin Robert B. , Manseau Eleanor J. , Goodwin Jane
Published: December 2022

Guinea pigs primed for cutaneous basophil hypersensitivity (CBH) with several soluble proteins or with sheep erythrocytes developed a systemic, delayed-onset, maculopapular rash when challenged parenterally with specific antigen. The rash was most readily induced 5 to 7 days after immunization, at a time when local CBH skin test reactivity was also optimal. Microscopically, the rash resembled local CBH skin test reactions, being comprised of a papillary dermal infiltrate of basophils and lymphocytes and a striking dilatation and compaction of superficial venules.

In addition to the systemic rash, animals expressing systemic CBH (SCBH) exhibited a striking eosinophilia at 24 hr which gave way to basophilia at 48 hr. Focal collections of eosinophils, and of smaller numbers of basophils, were found in the lungs and spleen; both eosinophils and basophils infiltrated the medulla of the thymus. Thus, basophilrich infiltrations are favored in the skin even after systemic challenge with antigen and occur only to a much smaller extent in other organs where eosinophils may predominate. These differences in the response of various organs to challenge with parenteral antigen suggest that as yet unidentified local factors play a determinative role in regulating the inflammatory response.

The pathogenesis of SCBH is not yet established, but it shares many of the properties of local CBH: histology, carrier specificity, development early after sensitization in the absence of detectable antibodies. Passive transfer has not been accomplished with serum alone but has been achieved irregularly with cells plus serum. SCBH may serve as a useful model for several disease states in man characterized by a systemic rash and eosinophilia, including certain types of drug reaction.
Cross-Reacting Antigens in Chemically Induced Sarcomas Are Fetal Determinants
Authors: Parker George A. , Rosenberg Steven A.
Published: December 2022

Serologic cross-reactivity has been demonstrated among three MCA-induced sarcomas in C57BL/6N female mice (MCA-2, MCA-3, and MCA-12) with a microcytotoxicity assay. Serum from mice bearing MCA-3 tumor was cytotoxic to both MCA-2 and MCA-3 tumor cells at a titer of 1:8. Sequential absorptions of this serum with syngeneic embryo cells completely eliminated cytotoxicity against MCA-2 cells without affecting the cytotoxic titer against MCA-3 cells. Serum hyperimmune to the MCA-3 tumor reacted with MCA-3, MCA-2, and MCA-12 tumors. Absorption of this serum with embryo cells eliminated cytotoxicity against MCA-2 and MCA-12 cells, but was incapable of lowering the titer against MCA-3 cells below 1:40. Similarly, serum hyperimmune to MCA-2 tumor was lytic to MCA-2, MCA-3, and MCA-12 before absorption, but was lytic only to MCA-2 cells after absorption with sygeneic embryo cells. Thus, the in vitro cross-reactivity between MCA-induced sarcomas is due to a common fetal antigen(s), which is distinct from the individual tumor-specific antigens of each tumor. Since these tumors do not exhibit cross-reactivity in in vivo challenge experiments, it appears that this fetal antigen is not responsible for in vivo immune protection.
Splenic Modifications Induced by Cyclophosphamide in C3H/He, Nude, and “B”-Mice
Authors: Kolb Jean-Pierre B. , Poupon Marie-France M. , Lespinats Geneviève M. , Sabolovic Domagoj , Loisillier Felix
Published: December 2022

The spleen cell population of adult C3H/He mice injected with a single sublethal dose of cyclophosphamide (CY) has been analyzed. An initial phase of spleen atrophy is followed by a considerable hypertrophy, and a progressive return to normal. During the phase of spleen atrophy, both B and T cell compartments are depleted, as estimated by the percentages of cells killed by anti-Thy 1–2 and anti-Ig antisera plus complement. During the stage of regeneration, the percentage of Ig + cells increases rapidly, and at the peak of splenomegaly, the percentage of Ig + cells is high whereas almost no Thy 1–2 + cells are detectable. Progressively, the spleen cell content returns to the original values. In thymodeprived mice (nude mice and B mice) the percentage of null cells increases during the stage of regeneration, and B mice develop a large number of Ig +-bearing cells.

Histologic examination shows that follicles (B-dependent areas) disappear 1 to 2 days before periarteriolar sheaths (T-dependent areas). At the peak of splenomegaly the architecture of the spleen is destroyed, and the interstitial tissue is composed of a dense and uniform layer of lymphoid cells. Progressively, the architecture returns to normal. In nude mice, the disappearance of follicles, and the appearance of a homogenous layer of lymphocytes has been observed.

When analyzed for their pattern of electrophoretic mobilities (E.M.), spleen cells from untreated mice reveal two peaks of E.M. 0.80 and 1.15 µm × s-1 × V-1 × cm-1. After CY treatment, during the step of splenic hypertrophy, these two peaks disappear, and a single peak of intermediate mobility appears. In T-deprived mice, a single peak of the same mobility is detected at this stage.

The nature and origin of cells which appear during the phase of regeneration are unclear, but their appearance in T-deprived mice argues against thymo dependence. These spleen cells have the ability to suppress the response of normal spleen lymphocytes to T and B cell mitogens.