bluuebunny/crossref_metadata_2025 huggingface dataset:
Alloantigen-induced cytotoxicity against syngeneic tumor cells: analysis at the clonal level.
Authors: Sensi M , Orosz C G , Bach F H
Published: December 2022

It has been shown that peritoneal exudate cells (PEC) from BALB/c mice immunized with minor histocompatibility antigens presented by DBA/2 or B10.D2 spleen cells are capable of lysing syngeneic YC8 tumor cells in a 4-hr 51Cr-release assay. In this study, we employed limiting dilution analysis to determine the frequency of CTL precursors (CTL-P) reactive against both the specific DBA/2 (or P815) target and the syngeneic tumor YC8. The mean frequency of anti-DBA/2 CTL-P in PEC from BALB/c mice immunized with DBA/2 was 1/302. Between one-third and one-fifth of limiting dilution microcultures that exhibited lytic activity against DBA/2 lymphoblasts (or P815) were also able to lyse YC8. No lysis of YC8 was observed in the absence of a parallel lysis on DBA/2 lymphoblasts or P815 target cells. T cell clones, derived by micromanipulation from microcultures selected for cytotoxic activity against YC8 and/or P815, maintained either the specific anti-allogeneic or the doubly reactive ( antiallogeneic plus anti-syngeneic tumor) phenotype. Fourteen clones (six specific and eight doubly reactive) were tested for cytotoxic activity on a panel of target cells with different haplotypes. All showed H-2-restricted specificity for minor histocompatibility antigens shared by DBA/2 and B10.D2. The restriction element for some of the clones mapped in the K region of the H-2 complex, whereas for other clones the restriction element mapped in the D region; both K- and D-restricted clones were able to lyse YC8. When the clones that exhibited lysis on YC8 were tested on two other BALB/c tumor targets, LSTRA, a Moloney virus induced lymphoma, and RL male-1, a radiation induced lymphoma, two of seven were found to lyse all three syngeneic tumor targets equally well, but not syngeneic BALB/c blasts. These clones were functionally categorized as conventional CTL because they were unable to proliferate when cultured with antigen in the absence of exogenous lymphokines, and were unable to produce lymphokine with IL 2 activity when stimulated by the appropriate splenocytes. When tested in vivo in a Winn assay, a strong anti-tumor activity against YC8 was exerted by the anti-DBA/2 clones DY4 -3 and DY16 -3. These clones lysed both YC8 and the immunizing target cells in vitro. No in vivo effect in neutralizing YC8 tumor growth was observed with clone D2-1, a clone that lysed DBA/2 targets but not YC8 in vitro.
Specific inhibition of in vitro lymphocyte transformation by an anti-pan T cell (gp67) ricin A chain immunotoxin.
Authors: Kernan N A , Knowles R W , Burns M J , Broxmeyer H E , Lu L , Lee H M , Kawahata R T , Scannon P J , Dupont B
Published: December 2022

The toxin A chain of ricin has been conjugated by a disulfide bond to a murine monoclonal antibody that recognizes the gp67kD antigen present on 95% of peripheral T lymphocytes. The immunotoxin retains both functions of its component parts: it binds to human peripheral blood lymphocytes, and it inhibits protein synthesis in a cellfree reticulocyte system. The immunotoxin has been evaluated for its ability to inhibit in vitro T lymphocyte transformation. In the presence of 20 mM NH4Cl, the immunotoxin decreases lymphocyte proliferation in response to phytohemagglutinin to less than 8% of untreated controls. The proliferative response in mixed lymphocyte culture and the development of allocytotoxic T cells is also dramatically inhibited by this immunotoxin. Monoclonal antibody alone does not inhibit these responses. Specificity of the immunotoxin has been established: the effect of the immunotoxin can be blocked by unconjugated monoclonal antibody, but not by a control monoclonal antibody that recognizes another T lymphocyte differentiation antigen or by a control monoclonal antibody that does not recognize human peripheral blood leukocytes. Treatment of human bone marrow cells with the immunotoxin preserves hematopoietic progenitor cells, as measured by granulocyte-macrophage, erythroid, and multipotential hematopoietic progenitor cell assays. These results indicate that an anti-pan T lymphocyte-ricin A chain immunotoxin is an effective agent against immunocompetent T lymphocytes in vitro, and may be an effective agent for use in clinical bone marrow transplantation.
Glucocorticosteroids stimulate polyclonal immunoglobulin production by cord blood mononuclear cells.
Authors: Orson F M , De Seau V , Pike S , Blaese M
Published: December 2022

Glucocorticosteroids (GCS) are in vitro polyclonal activators of immunoglobulin (Ig) production for human mononuclear cells (MC) from virtually all adult donors. However, GCS treatment of cord blood MC resulted in Ig production in only 12/41 samples. This GCS effect is T cell and monocyte dependent and is mediated in part by a soluble T cell replacing factor. The inconsistent response of cord MC could be due to either cellular or soluble factor differences from adults. In 11/12 samples tested, irradiated cord T cells did help adult B cells, but less than did irradiated allogeneic adult T cells. T cell suppression in cord samples is unlikely inasmuch as higher cord T cell numbers and proportions increased the number of responding cord samples. Cord monocytes function adequately, because monocytes supported GCS responses when cord non-T cells had sufficient T cell help. The T cell replacing factor was found in supernatants of unstimulated cord as well as in adult MC cultures, but was less than 50% as potent. Cord B cells did not develop GCS-induced Ig production with such supernatants, suggesting that cord B cells may require a higher concentration or more prolonged exposure to T cell help. With a 2:1 ratio of irradiated adult T cells to cord non-T cells, only 25% of cord samples responded to GCS (compared to greater than 95% of adult samples), but with a ratio of 4:1, 75% responded. IgM was the predominant isotype secreted by GCS-stimulated cord cells, but 6/14 samples also produced IgG and 8/14 produced IgA. Thus, the functional immaturity of both cord T and B cells exists for GCS-induced Ig production, but with appropriate conditions GCS can activate most samples of cord B cells to synthesize Ig.
Induction of suppressor T cells by interleukin 2.
Authors: Ting C C , Yang S S , Hargrove M E
Published: December 2022

The optimal concentration of interleukin 2 (IL 2) for maintaining the in vitro growth of T cells was quite different from that required for the induction of cytotoxic T lymphocytes (CTL) in nu/nu spleen cells. Higher concentrations of IL 2-containing preparations (10 to 30% v/v or 5 to 15 U/ml) were needed to promote the T cell growth, whereas lower concentrations (1 to 3% v/v or 0.5 to 1.5 U/ml) were needed to generate alloreactive CTL. It was further shown that the addition of high concentrations of IL 2 (10 to 30%) suppressed the generation of alloreactive CTL in conventional MLC. High concentrations of IL 2 induced the generation of antigen-nonspecific suppressor T cells in normal spleen cell cultures and augmented the generation of antigen-specific suppressor T cells in MLC. These suppressor cells suppressed the generation of CTL in fresh MLC and in polyclonal CTL cultures. These suppressor T cells could be induced by rat (spleen)-produced, murine (EL-4 cells) produced IL 2 preparations, and a purified human recombinant IL 2 (HR-IL2). The ability to induce suppressor cells correlated with the activity of IL 2 present in these preparations and was independent of their ability to induce cytotoxic effectors. These findings indicate that IL 2 may play a dual role in the regulation of CTL responses. We suggest that during antigen sensitization, the initial endogenous production of lower levels of IL 2 provided the second signal for the differentiation and proliferation of CTL. When higher levels of IL 2 were produced later, the suppressor T cell precursors were activated and differentiated into suppressor effectors to regulate the CTL response.
Noncovalent association of heavy and light chains in Rana catesbeiana immunoglobulins.
Authors: Mikoryak C A , Steiner L A
Published: December 2022

The unreduced immunoglobulins (Ig) in the bullfrog, Rana catesbeiana, dissociate into two components when subjected to electrophoresis or molecular sieving in dissociating solvents. One of these components is monomeric light chain and the other is a disulfide-bonded complex of heavy chains. This unusual behavior has been observed with all classes of bullfrog Ig that have been isolated and characterized previously: a high m.w. Ig that resembles mammalian IgM and two antigenically distinct varieties of low m.w. Ig. Light chains, isolated from the high m.w. Ig by gel filtration in 8 M urea, 1 M acidic acid, were found to contain, on average, 5.7 residues of half-cystine. None of these residues were in the free sulfhydryl form nor were they blocked by half-cystine. Moreover, none was alkylated after mild reduction of the high m.w. Ig. These findings indicate that none of the light chain half-cystine residues participate in an interchain disulfide bridge, and that most of the light chains contain three intrachain bridges. This unusual pattern of disulfide bonding appears to be responsible for the noncovalent association of heavy and light chains in this species.
Influence of the immunoglobulin heavy chain locus on expression of the VK1GAC light chain.
Authors: Fulton R J , Davie J M
Published: December 2022

The VK1GAC light chain represents the dominant V kappa structure employed in the antibody response of A/J mice to streptococcal group A carbohydrate ( GAC ). Two anti-idiotypic antisera, anti- Id5 and anti- Id20 , with specificity for the VK1GAC light chain were used to examine anti- GAC antibody responses in a series of inbred mouse strains that differ at the heavy chain constant region ( IgCH ) allotype locus. Both idiotypes were expressed in normal and immune sera from mice of most IgCH allotypes, except IgCHb (C57BL/6J) and IgCHf (CE/J). C57BL/6J mice expressed Id5 , but not Id20 , whereas CE/J mice did not express either idiotype. Testing of recombinant inbred strains between BALB/c and C57BL/6 indicated that the pattern of idiotype expression did not correlate with IgCH allotype. The C X B recombinants expressed all three idiotype patterns that were observed in the panel of inbred strains. Testing of allotype congenic mice between BALB/c and C57BL/6 showed that CB.20 and BC.8 mice were Id20 -, whereas BAB-14 mice were Id20 +, indicating that both VH and background (V kappa or regulatory) loci must be derived from BALB/c to obtain Id20 expression. The difference in the frequency of idiotype expression observed between BALB/c and BAB-14 mice indicates that the IgCH locus may exert a quantitative influence on the expression of this light chain. To examine the Id20 -, Id5 + antibodies of C57BL/6 mice, anti- GAC hybridomas were prepared. Of 16 C57BL/6-derived anti- GAC monoclonal antibodies, six were reactive with anti- Id5 and not with anti- Id20 . Isoelectric focusing of the purified kappa light chains from three of these antibodies revealed two distinct spectrotypes that co-migrated with the two known VK1GAC spectrotypes observed with A/J anti- GAC light chains. Idiotypic analysis of in vitro recombinants between the heavy and light chains of A/J and C57BL/6 monoclonal antibodies demonstrated that the C57BL/6 light chains were idiotypically similar to A/J light chains when they were free in solution or paired with A/J heavy chains. These results demonstrate that C57BL/6 mice can express a light chain that is very similar, if not identical, to the VK1GAC light chain, although the light chain is expressed in lower frequency and is paired with a distinct VH structure, which can mask expression of one of the VK1GAC idiotypes. These effects on V kappa expression map to at least three genetic loci: VH, CH, and an unlinked locus.
Synergism between lymphokines and muramyl dipeptide encapsulated in liposomes: in situ activation of macrophages and therapy of spontaneous cancer metastases.
Authors: Fidler I J , Schroit A J
Published: December 2022

Murine alveolar macrophages (AM) can be rendered tumoricidal after the i.v. injection of multilamellar vesicles (MLV) containing muramyl dipeptide (MDP) at a minimum dose of approximately 0.6 micrograms or less than a 10-fold dilution of cellfree culture supernatants from mitogen-stimulated F-344 rat lymphocytes rich in macrophage-activating factor (MAF) activity. Neither MAF at dilutions exceeding 1/10 nor MDP at doses lower than approximately 0.6 micrograms activated AM in situ to become tumor cytotoxic. The combination of these agents at subthreshold amounts (MAF 1/20, MDP 0.3 microgram), however, activated AM to significant levels of cytotoxicity. The synergism between MAF and MDP encapsulated in liposomes was also observed in therapeutic studies of mice bearing well established, spontaneous lung and lymph node metastases. Multiple i.v. injections of liposomes containing either 6.25 micrograms MDP or a 1/2 dilution of MAF resulted in the long-term survival (greater than 250 days) of 30% of the treated mice. Neither liposomes containing MAF at a 1/20 dilution nor MDP at doses lower than 0.6 microgram were therapeutically effective. Multiple injections of liposomes containing subthreshold amounts (MAF 1/20; and MDP 0.3 micrograms) of these agents, however, resulted in 50% survival in the treated group. These data allow us to conclude that the encapsulation of MAF and MDP within the same MLV is more efficient in inducing the destruction of metastases than either agent administered alone.
Immune serum against Anaplasma marginale initial bodies neutralizes infectivity for cattle.
Authors: Palmer G H , McGuire T C
Published: December 2022

The first demonstration of immune serum-mediated neutralization of Anaplasma marginale and identification of the involved surface antigens is presented. Anaplasma marginale initial bodies were purified from parasitized erythrocytes by using ultrasonic disruption and differential centrifugation. The initial bodies were morphologically intact, as determined by electron microscopy, were not agglutinated by anti-bovine erythrocyte sera, and were infective. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) demonstrated minimal erythrocytic contamination in the purified initial bodies. Rabbit antisera against initial bodies completely neutralized the infectivity of purified initial bodies for splenectomized calves, whereas normal rabbit serum did not alter infectivity. Nine initial body proteins were surface radioiodinated by using lactoperoxidase, and five of these were precipitated by the neutralizing antibody (105,000, 86,000, 61,000, 36,000, and 31,000 apparent m.w.). The results demonstrate that initial body determinants capable of inducing neutralizing antibody are present. The identification of surface radiolabeled proteins immunoprecipitated by the neutralizing antibody provides candidates to be tested as protective immunogens in cattle.
Polyclonal activation of the murine immune system by an antibody to IgD. VII. Demonstration of the role of nonantigen-specific T help in in vivo B cell activation.
Authors: Finkelman F D , Smith J , Villacreses N , Metcalf E S
Published: December 2022

Previous studies from this laboratory have shown that the injection of mice with an affinity-purified goat antibody to mouse IgD (GaM delta) induces T-independent polyclonal increases in: 1) B cell DNA synthesis, and 2) expression of surface Ia antigen and receptors for T helper factors, 1 to 2 days after injection. In addition, T-independent polyclonal increases in B cell number and IgG1 secretion are observed 6 to 7 days after injection. The administration of normal goat IgG (GIgG) along with GaM delta has been shown to augment GaM delta-induced polyclonal IgG1 secretion. To obtain information about the characteristics of the T help that is required for polyclonal antibody production in this model system, we investigated: 1) the length of the period during which GaM delta must be present to induce day 7 polyclonal antibody production, and 2) the kinetics of the induction of splenic T cell DNA synthesis. We found that GaM delta can be neutralized 3 days after injection by the administration of IgD without decreasing day 7 polyclonal IgG1 secretion, as long as mice are given GIgG at the time that GaM delta is neutralized. In contrast, polyclonal IgG1 secretion is greatly inhibited if GaM delta is neutralized 1 to 2 days after injection or if GaM delta is neutralized 3 days after injection, but GIgG is not administered at this time. Because GIgG can stimulate activated GIgG-specific T cells to secrete helper factors, but, unlike GaM delta cannot focus GIgG-specific T help polyclonally onto B cells, these findings suggest that nonspecific T help, rather than antigen-specific T help, is required in this system after day 3 for the induction of polyclonal IgG1 secretion. Determination of the kinetics of the induction of T cell DNA synthesis in this system by in vivo [3H] thymidine incorporation studies, as well as dual laser fluorescence-activated cell sorter analysis of T and B cell DNA content, indicate that T cells are induced to synthesize DNA 2 days after GaM delta injection and reach plateau rates of DNA synthesis 3 days after injection. Taken together, the GaM delta neutralization experiments and DNA synthesis studies suggest that one reason that GaM delta is required for 3 days in this system is to allow maximal activation of GIgG-specific T cells, which when stimulated later by GIgG secrete nonantigen-specific helper factors that induce GaM delta-activated B cells to secrete IgG1.
Endogenous blood levels of corticosterone control the immunologic cell mass and B cell activity in mice.
Authors: del Rey A , Besedovsky H , Sorkin E
Published: December 2022

An inverse relation between endogenous levels of glucocorticoids and splenic mass and cellularity was detected. Decreased endogenous corticosterone blood levels result in an increased number of immunoglobulin-secreting cells in the spleen, a fact that cannot be attributed only to the expansion of the lymphoid cell mass. The opposite phenomenon was observed in animals with high corticosterone blood levels, which showed reduced numbers of immunoglobulin-secreting cells. It is concluded that endogenous glucocorticoid levels contribute to the control of B cell activity and possibly to the interaction of these cells with other immunologic cells.
Interactions between T lymphocyte subsets supported by interleukin 2-rich lymphokines produce acute rejection of vascularized cardiac allografts in T cell deprived rats.
Authors: Heidecke C D , Kupiec-Weglinski J W , Lear P A , Abbud-Filho M , Araujo J L , Araneda D , Strom T B , Tilney N L
Published: December 2022

A series of adoptive transfer studies were performed to define both the role of lymphocyte subpopulations and of lymphokines in the reestablishment of immunologic responsiveness towards vascularized organ allografts in T cell-deprived B rats. Acute rejection of otherwise indefinitely surviving cardiac allografts (10.9 +/- 2.9 days) occurred when sensitized T cells were adoptively transferred with a course of partially purified, lectin-free IL 2-conditioned medium (IL 2CM), whereas rats receiving T cells alone rejected their grafts at 19.8 +/- 4.6 days (p = 0.025). Rejection is mediated primarily by alloactivated T helper cells highly enriched for W3/25 phenotype. The relationship between the mean allograft survival time and the cell number transferred was of exponential order. In contrast to rejection occurring after the transfer of intact T cells, that produced by W3/25+ cells was independent of the administration of exogenous IL 2CM. W3/25+ T helper cells were also found to be less potent in mediating graft rejection than were whole T cells when limited cell numbers were transferred. The OX8+ T cytotoxic/suppressor cell subpopulation, even when supplied with a course of IL 2CM, was unable to bring about rejection, but induced transient graft enlargement in about 50% of the animals. Recombination of individual subsets caused acute rejection at a rate comparable to that of unseparated T cells when IL 2CM was supplied (10.8 +/- 1.0 days), in contrast to those rats which received the recombined inoculum in the absence of IL 2CM (16.8 +/- 2.2 days, p = 0.025). Lymphocyte migration studies revealed a more vigorous homing of OX8+ cells to the allograft as compared to W3/25+ cells. However, to produce maximal accumulation of transferred cells in the graft, both T cell subsets supplemented by lymphokines must be introduced together into T cell-deprived hosts.
An analysis of functional T cell recognition sites on I-E molecules.
Authors: Needleman B W , Pierres M , Devaux C A , Dwyer P N , Finnegan A , Sachs D H , Hodes R J
Published: December 2022

The recognition of I-E molecules by antigen-specific T cells was studied to determine if one or multiple topographic sites on the I-E molecules can function as restricting elements for T cells. A panel of 14 I-Ek-specific monoclonal antibodies (mAb) was used to inhibit T cell proliferation induced by antigens, the recognition of which was restricted by I-E-encoded determinants. These antibodies gave patterns of inhibition that were similar for three long-term antigen-specific T cell lines. Multiple distinct patterns of inhibition, however, were observed when a series of antigen-specific I-E-restricted T cell clones was studied. Differences were identified even among clones expressing apparently similar antigen specificities and MHC restriction. The observed inhibition by these antibodies appeared to be caused by specific steric or allosteric interference with T cell recognition of antigen and Ia. Based on the differences in patterns of inhibition, it was possible to infer the existence of distinct sites or conformations on the I-E molecule that are functionally involved in antigen-specific T cell recognition.
Isolation and characterization of an I-A-restricted T cell clone with dual specificity for poly(Glu60Ala30Tyr10) (GAT) and Mlsa,dl.
Authors: Jenkins M K , Melvold R W , Miller S D
Published: December 2022

We have isolated a BALB/c (H-2d, Mlsb) T cell clone (JTL-G12) specific for the synthetic polypeptide antigen poly(Glu60Ala30Tyr10) (GAT) in the context of self I-A determinants and for Mlsa,d antigens in the absence of GAT. JTL-G12 proliferation in response to GAT was mapped to the Kd, I-Ad subregions by using inbred H-2 congenic and recombinant strains. In addition, monoclonal antibody directed against I-Ad but not Kd or I-As determinants blocked JTL-G12 proliferation in response to GAT presented by syngeneic splenocytes, indicating I-A restriction. The Mls cross-reactivity of this clone was verified by using a panel of inbred strains bearing the Mlsa,b,c,d alleles and by using BXD recombinant inbred strains bearing the Mlsa allele or the Mlsb allele. All of the Mlsa BXD strains of the H-2d or H-2b haplotypes stimulated JTL-G12 in the absence of GAT, whereas all of the Mlsb BXD strains were nonstimulatory. This response pattern is in complete accordance with recognition of the Mlsa determinant encoded by Mls or closely linked loci on chromosome 1. JTL-G12 proliferation in response to GAT/I-Ad and Mlsa,d determinants could be blocked with a monoclonal antibody (GK1.5) directed against L3T4, a structure involved in class II major histocompatibility complex antigen recognition. These results suggest that antigen/class II responsiveness, Mls reactivity, and expression of L3T4 can be properties of a single T cell population.
An immunofluorescence analysis of the ontogeny of myeloid, T, and B lineage cells in mouse hemopoietic tissues.
Authors: Velardi A , Cooper M D
Published: December 2022

The population dynamics of granulopoietic cells, B-lineage cells, and T lymphocytes were analyzed by immunofluorescence in mouse hemopoietic tissues as a function of age. Mac-1+ myeloid cells were present on day 11 of gestation in the liver, where they peaked shortly after birth and declined subsequently. Waves of myeloid population growth began in spleen and bone marrow by days 15 and 19, respectively. Mac-1+ cells increased in number to relatively low plateau levels in spleen by the 3rd wk after birth, whereas in the bone marrow higher plateau levels were reached around 3 mo of age. The 14.8 monoclonal antibody was utilized as one marker of B-lineage precursor cells. 14.8+ cells were detected in the liver on day 11 of gestation, reached peak numbers during the first week after birth and decreased thereafter. On day 15 and 19, 14.8+ cells were found in spleen and bone marrow, respectively, and progressively increased in numbers to reach plateau levels in both sites by 3 mo of age. Mu+ pre-B cells appeared in significant numbers in the 13-day fetal liver, reached a peak shortly after birth, and disappeared from the liver by the end of the second postnatal week. Pre-B cells were found in the spleen and bone marrow on days 15 and 19, respectively. In the spleen pre-B cells reached peak values at birth and disappeared 2 wk later. In spite of the sequential appearance of mu+ pre-B cells in fetal liver, spleen, and bone marrow, their sIgM+ B cell progeny appeared in all these hemopoietic tissues on day 17 of gestation. In the liver, sIgM+ B cells reached their peak at birth and declined thereafter. In the spleen and bone marrow, B cells increased to plateau levels between 1 and 4 mo of age. Thy-1.2+ T cells were relatively late acquisitions in all three hemopoietic tissues. Finally, the expression of the 14.8 antigen by mu+ cells was examined as a function of gestational age. While pre-B cells from day-13 fetuses had no detectable 14.8 antigen, the antigen was weakly expressed on the vast majority of the mu+ pre-B cells by day 17 of gestation. Newborn liver cells expressing 14.8 antigen were found to include a small proportion of cells with peroxidase+ granules. Thus, demonstration of rearrangement and expression of immunoglobulin genes may be required for precise identification of cells of B lineage early in ontogeny.
Relationship between immune system and gram-negative bacteria. II. Natural killer cytotoxicity of Salmonella minnesota Rb 345-unbound human peripheral blood lymphocytes.
Authors: Antonaci S , Jirillo E , Ventura M T , Michalek S M , Bonomo L , McGhee J R
Published: December 2022

Spontaneous binding of human peripheral blood lymphocytes (PBL) to bacteria represents a promising approach for the characterization of lymphocyte subsets mediating different functions. In the light of previous findings on the high degree of spontaneous adherence of S. minnesota Rb cells to PBL, we have evaluated the natural killer (NK) cytotoxicity of PBL subpopulations that fail to bind to Rb bacteria. The S. minnesota Rb-unbound cell fraction exhibits higher levels of cytotoxic capacity, which is related to a more elevated frequency of active NK cells, as determined in an agarose-single cell cytotoxic assay. Moreover, the cytotoxic activity of the unbound fraction is additionally boosted by interferon-alpha pretreatment. The effector cells bear Fc gamma receptors that are involved in NK cell lysis, because a decrease of NK activity is observed after immune complex modulation of the receptors. Finally, these cells, which display a high percentage (approximately 70%) of typical large granular lymphocyte morphology, express HNK-1, T10, T8, and M1 antigens, and to a lesser extent T3 and T4 antigens. These data indicate a selective enrichment of NK cells in the S. minnesota Rb-unbound fraction.
Measles virus-specific T4+ human cytotoxic T cell clones are restricted by class II HLA antigens.
Authors: Jacobson S , Richert J R , Biddison W E , Satinsky A , Hartzman R J , McFarland H F
Published: December 2022

We have generated measles virus-specific T cell clones from a patient with multiple sclerosis who has been described previously as a strong responder to measles virus. T cell clones were screened on the basis of their capacity to proliferate to measles virus. The cell surface phenotype of each clone was OKT3+, OKT4+, OKT8-. The majority of these clones (11 of 14) were cytotoxic for their autologous measles virus-infected lymphoblastoid B cell target. This cytotoxicity was specific for measles virus inasmuch as these T cell clones could not lyse influenza or mumps virus-infected B cell targets. By using a panel of HLA-defined measles virus-infected B cell lines, these clones were shown to recognize measles virus in the context of HLA class II determinants. A monoclonal antibody that recognizes HLA class II monomorphic determinants (L243), but not a monoclonal antibody that recognizes HLA class I antigens (W6/32), inhibited the lysis of the autologous measles virus-infected B cell target by these T cell clones. These results demonstrate that these cytotoxic T cell clones specific for measles virus are HLA class II restricted.
Interferon-gamma reduces macrophage-suppressive activity by inhibiting prostaglandin E2 release and inducing interleukin 1 production.
Authors: Boraschi D , Censini S , Tagliabue A
Published: December 2022

Normal peritoneal M phi of C3H/HeN mice were able to suppress lymphocyte proliferation in a dose-dependent fashion when added to Con A-pulsed spleen cell cultures. However, M phi-suppressive activity could be partially or completely reduced by in vitro pre-exposure to nonimmune IFN-alpha or immune recombinant IFN-gamma. For both IFN-alpha and IFN-gamma, reduction of M phi suppression was marginal at 10(1) U/ml and became highly significant at 10(2) to 10(3)/ml. The ability of IFN-alpha and IFN-gamma to modulate M phi suppression appears to be related to distinct mechanisms. In fact, impairment of M phi suppression by IFN-alpha occurred in parallel to the decrease of M phi capacity to produce PGE2 and the oxygen intermediate O2-, two molecules responsible for M phi-suppressive activity. In contrast, M phi exposed to IFN-gamma showed only impairment of PGE2 production, whereas O2- release was not significantly affected. Furthermore, at variance with IFN-alpha, IFN-gamma directly stimulated M phi to synthesize and release IL 1, a monokine known to promote lymphocyte proliferation.
Structural analysis of an HLA-B7 antigen variant detected by cytotoxic T lymphocytes.
Authors: Taketani S , Krangel M S , Spits H , de Vries J , Strominger J L
Published: December 2022

It has been demonstrated previously that lymphocytes of donor CF (HLA-A29,w33; B7,14) are not recognized by the HLA-B7-specific CTL clone HG-31. This report presents a structural comparison of the HLA-B7 antigen of donor CF with a "normal" HLA-B7 antigen, derived from the cell line JY. Isoelectric focusing showed that CF HLA-B7 heavy chains were more acidic than JY HLA-B7 heavy chains by the equivalent of a single charge. High pressure liquid chromatography and ion exchange chromatography comparisons of double-labeled tryptic peptides revealed a single detectable difference, which corresponded to the tryptic peptide spanning residues 112 to 121 on the HLA-B7 heavy chain. Although the complete amino acid sequence of this peptide was not obtained, the partial sequence indicates a substitution of an unidentified amino acid for tyrosine at position 116 of the heavy chain. This residue is found to vary among HLA specificities and to be altered in many H-2Kb mutants.
Preparation, isolation, and immunochemical studies of the cyanogen bromide peptides from a retinal photoreceptor cell autoantigen, S-antigen.
Authors: Gregerson D S , Putterman G J
Published: December 2022

Purified bovine retinal S-antigen (50,000 m.w.) was treated with cyanogen bromide, producing seven major and several minor fragments. Six of the major and one of the minor components were isolated by reverse phase high performance liquid chromatography. The peptides were characterized with respect to size by urea-SDS-gel electrophoresis, by amino acid composition, and by their ability to bind antibodies, raised in rabbits immunized with purified bovine S-antigen, in both competition and direct enzyme-linked immunosorbent assays. Four of the purified peptides were found, by the direct assay, to bind antibodies in immune sera raised to the intact antigen. Peptides that were negative, or only weakly bound, in the direct enzyme immunoassay were subsequently conjugated to a carrier, poly-L-Glu-Ala-Tyr, and were retested in the enzyme immunoassay in which a peptide of about 25 residues was also found to contain an antigenic determinant. The same five peptides were positive in the competition assays. Isolation of the peptides and gel electrophoresis under reducing and nonreducing conditions revealed that two of the peptides in the reaction mixture were joined by a disulfide linkage.
Antigens of Dirofilaria immitis which are immunogenic in the canine host: detection by immuno-staining of protein blots with the antibodies of occult dogs.
Authors: Boto W M , Powers K G , Levy D A
Published: December 2022

We describe here the profiles of antigens of Dirofilaria immitis that are immunogenic in dogs. Purebred beagle dogs were inoculated with a standard number of the infective third-stage larvae of the heartworm. Microfilaremia and anti-heartworm antibody levels were monitored by the Knott's test and by ELISA, respectively. Antibody-binding polypeptides were detected by immunoperoxidase staining of protein blots of detergent extracts of adult parasites from SDS-PAGE gels. Densitometric scans of these blots revealed considerable variation in the profiles of dirofilarial antigens detected by the sera obtained at different stages of the infection. Sera obtained during the prepatent phase, i.e., 3 mo post-infection (titer 1:2000), detected antigens of Mr 75, 80, 100, 130, and 200 kilodaltons. In addition to these antigens, sera obtained at the onset of microfilaremia, 6 mo post-infection (titer 1:30,000), detected antigens of Mr 15, 16, 18, 34, and 38 kilodaltons. Protein blots stained with sera from dogs, which subsequently cleared the microfilaremia and in which the infection became occult, showed that most of the antibody reactivities were directed at antigens of Mr 15, 21, and 38 kilodaltons. The variations in the species of molecules detected in sera obtained during the course of an infection seem to reflect fluctuations in the levels of specific antibodies directed at the individual heartworm antigens. We discuss the apparent role that the metamorphic differentiation of the parasite may play in determining the levels of reactivities of these antibodies at specific stages of the infection.
Homologous and distinctive antigens of Onchocerca volvulus and Dirofilaria immitis: detection by an enzyme-linked immuno-inhibition assay.
Authors: Boto W M , D'Antonio R , Levy D A
Published: December 2022

The limited accessibility of specimens of Onchocerca volvulus has hampered investigations of factors that determine the clinical course of infection of man by this filarial nematode. Parasites antigenically related to O. volvulus have been variously proposed as alternative sources of test antigens. The present study has utilized serologic methods to probe the degree of identity of the antigenic constituents of O. volvulus and Dirofilaria immitis, the dog heartworm. 35S-methionine-labeled proteins of dog heartworm microfilariae (mf) were analyzed in immunoprecipitation assays by using a panel of sera drawn from humans diagnosed for onchocerciasis and from canines infected with D. immitis. Unique and common antigens detected by these sera were resolved in polyacrylamide gels. The majority of high-titered human and infected dog sera reacted with antigenic molecules estimated at 15, 16, 28, 42, 54, 66, and 100 kilodaltons. In addition, a subset of these sera detected large amounts of a 75 kilodalton antigen. Human anti-O. volvulus antibodies were found to bind to intact glutaraldehyde-fixed mf of D. immitis in immunoperoxidase assays. In enzyme-linked immunosorbent assays, the same sera reacted with extracts of three distinct developmental stages of the canine pathogen--namely, the mf, embryonated eggs, and adult worms. Preincubation of dirofilarial antigens with human anti-O. volvulus sera inhibited the binding of dog anti-heartworm serum by as much as 70%. Sera of guinea pigs hyperimmunized with mf of O. volvulus inhibited the binding of the immune dog serum to the same extent. These results represent evidence for extensive homology in the antigenic composition of O. volvulus and D. immitis.
Delayed hypersensitivity granuloma formation around Schistosoma mansoni eggs in vitro. III. Granuloma formation and modulation in human Schistosomiasis mansoni.
Authors: Doughty B L , Ottesen E A , Nash T E , Phillips S M
Published: December 2022

An in vitro model of granuloma formation was used to study the cellular immune responses of Schistosoma mansoni-infected patients. The purposes of this study were to determine the relationship of granulomatous hypersensitivity to S. mansoni eggs in recent, well-defined infections and long-term chronic infections, and to determine the role of T cell subsets (OKT3, 4, and 8) defined by monoclonal antibodies in granulomatous hypersensitivity. Peripheral blood mononuclear cells obtained from patients with recent S. mansoni infections demonstrated increased granulomatous hypersensitivity responses in vitro when compared to peripheral blood mononuclear cells obtained from patients infected for 5 yr or more. The selective removal of infected for 5 yr or more. The selective removal of OKT3+ or OKT4+ cells reduced the ability of peripheral blood mononuclear cells to form granulomas in vitro. Positive selection for OKT4+ T cells produced optimal granulomatous hypersensitivity when compared to that produced by the unfractionated peripheral blood mononuclear cell population. OKT8+ cells demonstrated no ability to form granulomas in vitro. Selective removal of OKT8+ T cells produced variable results in the ability of the remaining peripheral blood mononuclear cells to form granulomas in vitro. These studies demonstrate the feasibility of investigating granulomatous hypersensitivity and immunoregulatory mechanisms operative in S. mansoni-infected patients by using in vitro technology.
Intrathymic lymphopoiesis: stromal cell-associated proliferation of T cells is independent of lymphocyte genotype.
Authors: Kyewski B A , Travis M , Kaplan H S
Published: December 2022

We analyzed the genetic restriction of direct cell-cell interactions between thymocytes and a) cortical epithelial cells, b) macrophages, and c) medullary dendritic cells in the mouse thymus. Thymectomized (C3H X C57BL/Ka)F1 hybrid mice were doubly grafted with P1 and P2 neonatal thymus grafts, were lethally irradiated, and were reconstituted with a mixture of P1 and P2 bone marrow cells which differed in the Thy-1 locus. The contributions of both parental inocula to the composition of the free and stromal cell-associated T cell compartments were analyzed separately in thymic grafts of each parental strain. The lymphoid composition in both compartments essentially reflected the peripheral T cell-chimerism in the host. The development of lymphostromal complexes was not restricted by the genotype of the partner cells. Statistical analysis of the distributions of P1 and P2 T cells among free thymocytes and within individual lymphostromal complexes, however, suggests that the T cells of an individual complex are the progeny of oligoclonal proliferation. Thus, both epithelial cells and bone marrow-derived stromal cells seem to be involved in different stages of intrathymic lymphopoiesis.
Homologous monoclonal antibodies induce idiotope-specific suppression in neonates through maternal influence and in adults exposed during fetal and neonatal life.
Authors: Rothstein T L , Vastola A P
Published: December 2022

The fine specificity of idiotype suppression induced early in ontogeny was investigated in the murine A/J anti-azophenylarsonate (Ar) response. Suppression was induced with two hapten-inhibitable, homologous monoclonal anti-idiotopic antibodies, AI and MB, that recognize partially overlapping sets of Ar-immune antibodies. Suppression was found to be idiotope-specific when adult mice were exposed to anti-idiotope as neonates; suppression was also idiotope specific when adult mice were exposed to anti-idiotope during fetal (through maternal inoculation) and neonatal life. Of particular interest, anti-idiotope, administered maternally, induced suppression in offspring first immunized with Ar as neonates, and this suppression was idiotope specific too. Thus, AI and MB induce idiotope-specific suppression in mice exposed to anti-idiotope early in ontogeny. These results parallel previous findings in adult mice and suggest that the mechanism of suppression in very young mice is the same as that in adults.
A comparison of the stimulatory activities of lymphoid dendritic cells and macrophages in T proliferative responses to various antigens.
Authors: Guidos C , Wong M , Lee K C
Published: December 2022

The identities of murine accessory cells and the mechanism by which they process antigen and stimulate T cell proliferation have been examined with cell separation techniques and specific agents to block antigen catabolism. Using preparations of splenic dendritic cells (DC) and macrophages (M phi) with minimal cross-contamination, we found that only DC could induce syngeneic mixed leukocyte reaction (MLR), whereas both DC and M phi could initiate allogeneic MLR. This observation may have significant implications for syngeneic MLR as a manifestation of self Ia recognition, and for the cell type that defines self Ia during ontogeny. DC and M phi could present soluble antigens such as purified protein derivative of tuberculin (PPD) and Salmonella flagellin about equally well to antigen-specific T cell lines. M phi, however, were much more effective than the non-phagocytic DC at inducing T cell proliferation to whole Corynebacterium parvum organisms. These differences could not be attributed to differences in antigen uptake. The results suggest that the bacteria must be ingested and processed by phagocytes before T cell activation. Using the lysosomotropic agent chloroquine to inhibit antigen catabolism in accessory cells, we found that the presentation of large antigens by M phi and DC was abolished by chloroquine treatment, whereas T cell activation by antigens (such as PPD or integral membrane Ia for MLR) that apparently required no processing was relatively insensitive to chloroquine. Thus, in addition to differences between cells, discrete functions within each cell type can also be distinguished.
Ontogeny of B cell markers in the human fetal liver.
Authors: Hofman F M , Danilovs J , Husmann L , Taylor C R
Published: December 2022

Human fetal liver was examined during various stages of gestation for the presence of B cells by using immunoglobulin isotype markers and monoclonal B cell antibodies. Frozen sections were studied with the use of single and double staining methods. The B cell monoclonal antibodies used were BA1, which defines both mature and immature B cells; B1, which identifies mature B cells; and B532, which binds to activated mature B cells. The data indicate that both BA1 and mu+ cells are present at 12 wk gestation, and increase in frequency with age. Delta and B1-bearing cells are detected only later in fetal life. Phenotypically identifiable T cells are present at low frequencies in the fetal liver throughout the time period examined (12 to 21 wk). At 12 to 13 wk gestation, the numbers of kappa- and lambda-chain-positive cells are two to three times greater than the number of mu+ cells. Based on morphology and staining with OKM1, these light chain-bearing cells appear to be non-lymphoid, most likely cells of macrophage origin that have phagocytosed maternal IgG. Our results show that the monoclonal antibodies reacting with subsets of B cells in adults can also be used to define distinct subsets of B and pre-B cells in the fetal liver.
Ontogeny of B lymphocyte function. XV. A role for thymus cells in the maturation of the capacity of a subpopulation of B cells to re-express surface immunoglobulin after treatment with anti-immunoglobulin.
Authors: Chen Y W , Jacobson E B , Siskind G W
Published: December 2022

The maturation of the C57BL/6 B cell population to be able to re-express surface immunoglobulin (sIg) after its removal by treatment with rabbit antimouse Ig (RAMIg) was studied in a cell transfer system. It was found that thymus cells were required for the maturation of a subset of the B cell population to be able to re-express sIg. The B cell population of irradiated, thymectomized mice reconstituted with spleen cells from donors under 2 wk of age remained deficient in their ability to re-express sIg even after 4 wk residence in the cell transfer recipient. In contrast, if adult thymus cells were transferred together with the immature B cells, the B cell population matured to be able to re-express sIg after treatment with RAMIg. Approximately one-third of the B cell population appears to require thymus cells for this maturation. The maturation of the thymus cell population to be capable of mediating this maturation of the B cell population occurs in two steps: between 2 and 3 and between 3 and 4 wk of age. This timing corresponds to the age at which the B cell population of C57BL/6 mice normally acquires the capacity to re-express sIg, which we have previously shown to also occur in two steps. Thymus cells from 3-wk-old donors can mediate the first step in B cell maturation to be able to re-express sIg, but cannot mediate the second step in this maturation of the B cell population. Thymus cells from 4-wk-old donors can mediate both steps in the maturation of the B cell population. The results suggest that thymus cells are involved in regulating some aspects of B cell differentiation.
Localization of human myeloid-associated surface antigen detected by a panel of 20 monoclonal antibodies to the q12-qter region of chromosome 11.
Authors: van Kessel A G , Tetteroo P , van Agthoven T , Paulussen R , van Dongen J , Hagemeijer A , von dem Borne A
Published: December 2022

Human x mouse myeloid cell hybrids were obtained after fusion of human leucocytes and the murine myeloid cell line WEHI-TG. The hybrids were tested for reactivity with a panel of monoclonal antibodies with a known myelocytic, monocytic, or myelomonocytic specificity. Twenty antibodies, all of which bind specifically to the surface of human myeloid cells, exhibited very similar reactivity patterns with the hybrid clones. Chromosomal analysis of hybrid cell metaphases revealed that the gene(s) involved in the expression of the antigen(s) recognized by these antibodies must be located on human chromosome 11 in the region q12-qter. These results are compatible with the evidence obtained from other studies that several, if not all, of the myeloid-specific antibodies used are reactive with a similar antigenic determinant present on human myeloid cells.