bluuebunny/crossref_metadata_2025 huggingface dataset:
Specificity of H-2-linked Ir gene control in mice: demonstration of T helper cells recognizing branched synthetic polypeptides in low responder mice.
Authors: Stötter H , Imm A , Meyer-Delius M , Rüde E
Published: December 2022

This report provides evidence for the presence of T helper cells capable of recognizing the polypeptide antigens T6-A--L and (H,G)-A--L in low responder mice of H-2k and H-2b haplotypes, respectively. Mice were primed in vivo with the T6-A--L-avidin-(H,G)-A--L complex or, in the case of T6-A--L in H-2k mice, with the cross-reactive and permissive antigen T6-S--L. T helper cells cooperating with DNP-primed B cells could be rechallenged in vitro with the DNP-conjugates of T6--A--L or (H,G)-A--L, although the cells were of low responder type with respect to these antigens. This implies that T cell-macrophage interaction required for restimulation is apparently not defective in these low responders. The implications of these results for the concept of Ir gene control are discussed.
Suppression of phagocytosis and chemotaxis by cell wall components of Staphylococcus aureus.
Authors: Musher D M , Verbrugh H A , Verhoef J
Published: December 2022

The ability of S. aureus to resist phagocytosis by polymorphonuclear leukocytes (PMN) is thought to be an important virulence factor for this microorganism. We have studied the effect of 3 major cell wall components of S. aureus, peptidoglycan, protein A, and teichoic acid, on PMN function. Phagocytosis and chemotaxis were both inhibited by prior incubation of PMN with peptidoglycan. This effect was dose- and time-related; incubation with as little as 2.5 micrograms/ml for 30 min produced a discernible suppressive effect. Suppression of PMN function was independent of the presence of human serum but was abolished by rabbit antiserum to peptidoglycan. Addition of peptidoglycan to PMN stimulated a prolonged chemiluminescence response that was greater when the peptidoglycan was first incubated in normal serum, perhaps reflection opsonization of this particulate material. Although protein A also suppressed phagocytic and chemotactic capabilities of PMN, this effect was observed only in the presence of serum and was eliminated by absorbing Ig. Centrifugation of the serum + protein A mixture showed that the suppressive effect was contained in the precipitated sediment. Complexes of Ig alone were not suppressive. Teichoic acid in concentrations less than or equal to 100 micrograms/ml had no adverse effect on PMN function. These studies describe ways in which peptidoglycan and protein A may interfere with phagocytosis and chemotaxis of human PMN, thus giving evidence for the role of these cell wall components as virulence factors.
T cell-replacing factor- (TRF) induced IgG secretion in a human B blastoid cell line and demonstration of acceptors for TRF.
Authors: Muraguchi A , Kishimoto T , Miki Y , Kuritani T , Kaieda T , Yoshizaki K , Yamamura Y
Published: December 2022

IgG-secretion was induced in a human B blastoid cell line, CESS, by the addition of partially purified T cell-derived helper factor(s) (TRF), which had been obtained from PHA-stimulated human T cells. The number of IgG-producing cells in CESS cells reached its maximal level (10% of total cells) within 48 hr after the addition of TRF. TRF did not affect the proliferation of CESS cells and the block of cell proliferation with hydroxyurea did not inhibit the increase of IgG-producing cells, showing that TRF induced IgG-production in CESS cells without any requirement of cell division. TRF activity was completely removed by CESS cells but TCGF-activity in the same preparation was not absorbed with CESS cells. On the other hand, TCGF-dependent human killer cells absorbed TCGF activity but not TRF activity in the same preparation. The binding of 125I-labeled factor(s) on CESS cells was also demonstrated. These results showed the presence of acceptors for TRF on the surface of CESS cells and this cell line will provide useful means for the chemical characterization of acceptors and for the study of the mechanisms of the signal transmission through acceptors.
Proteolysis of the monomeric and dimeric C5b-9 complexes of complement: alteration in the susceptibility to proteases of the C9 subunits associated with C5b-9 dimerization.
Authors: Yamamoto K , Migita S
Published: December 2022

The C5b-9 monomer having the sedimentation coefficient of 23S was extracted from the rabbit erythrocyte membranes that had been treated with a limiting amount of C9-deficient human serum and of 125I-C9. Upon proteolysis by trypsin and chymotrypsin, the C9 subunits of this complex were cleaved by these enzymes at multiple sites, yielding fragments with m.w. ranging fro 40,000 to 19,000. The uncomplexed C9 was also cleaved by both enzymes at multiple sites. By contrast, the C9 subunits of the C5b-9 dimer were found to be totally insusceptible to chymotrypsin under the conditions studied (37 degrees C; 24 hr) and only partially susceptible to trypsin (33% of the C9 subunits were cleaved by trypsin into 2 fragments during incubation at 37 degrees C for up to 24 hr). Therefore, these results indicate that, although the binding of C9 molecules to the C5b-8 complex (C5b-9 monomer formation) does not significantly affect the susceptibility to proteases of the C9 molecules, C5b-9 dimer formation markedly limits the accessibility of proteases to the C9 subunit molecules. A implication of this finding to a role for C9 in C5b-9 dimerization is discussed.
Lymphocyte collaboration is required for induction of murine monocyte procoagulant activity by immune complexes.
Authors: Schwartz B S , Edgington T S
Published: December 2022

Both aggregated IgG and soluble immune complexes composed of human serum albumin and specific antibody at a 4:1 ratio were demonstrated to rapidly induce an increase in procoagulant activity (PCA) of murine peripheral blood mononuclear cells. Direct cytologic assays as well as fractionation of cells by adherence to plastic established the monocyte as the cell producing PCA, and demonstrated further that almost all monocytes, rather than a subpopulation, participated in the response. Monocytes that were depleted of lymphocytes did not respond to these stimuli with production of PCA; that is, induction of monocyte PCA was observed only when stimulation was performed in the presence of lymphocytes. Induction was equally efficient when only the lymphocyte was stimulated by aggregated IgG or soluble immune complexes, washed free of unbound stimulus, and added to monocytes. This suggests a unidirectional pathway by which the stimulus triggers lymphocytes after which there is direct lymphocyte-mediated induction of monocytes to produce PCA. Evidence for participation of soluble lymphocyte products was not obtained, and it is suggested that this series of events may be mediated by direct lymphocyte contact with responding monocytes. The role of this pathway in the genesis of immunologically initiated tissue lesions is discussed.
Characterization and isolation of IgE class-specific suppressor factor (IgE-TsF) I. The presence of the binding site(s) for IgE and of the H-2 gene products in IgE-TsF.
Authors: Suemura M , Shiho O , Deguchi H , Yamamura Y , Böttcher I , Kishimoto T
Published: December 2022

IgE-specific reverse plaque assay for the direct comparison of the IgE and IgG antibody responses was established and the method was employed for the assessment of the activity of IgE class-specific suppressor factor (IgE-TsF). In the in vitro culture system, the addition of IgE-TsF to DNP-KLH-primed spleen cells inhibited an antigen-induced increase of IgE-producing cells but did not show any effect on the IgG or IgM responses. Absorption of IgE-TsF with IgE-producing hybridoma cells removed the suppressor activity but IgM-producing hybridoma cells did not absorb the suppressor activity. The suppressor activity of IgE-TsF was removed by murine IgE-conjugated Sepharose column but not by IgM-, IgG-, or human IgE-conjugated column. The suppressor activity was eluted from IgE-column with glycine-HCI buffer, pH 3.2, or acetate buffer, pH 4.0, and the suppressor factor eluted from IgE-column was reabsorbed by anti-H-2d conjugated column. The results showed that IgE-specific suppressor factor was composed of the binding sites for IgE molecules and the H-2 gene products.
Anti-immunoglobulin antibodies. II. Expression of individual and cross-reactive idiotypes on syngeneic and homologous anti-idiotype antibodies.
Authors: Bona C , Kearney J
Published: December 2022

Syngeneic anti-(anti-Id) antibodies were prepared against BALB/c anti-A48Id antibodies, BALB/c anti-460Id monoclonal antibodies, and A/J anti-J558 IdI monoclonal antibodies. With these anti-(anti-Id) antibodies we identified cross-reactive idiotypes on syngeneic and homologous anti-A48Id and anti-460Id antibodies. By contrast, tbe idiotypic determinants of A/J anti-J558 IdI monoclonal antibodies were not shared by other syngeneic, homologous, or xenogenic anti-J558 IdI or IdX antibodies. These results suggest that idiotype-antiidiotype reactions that serve as regulatory controls within the immune system are characteristic for each particular antigen system, strain, or species and that such interactions make the system self-limited with respect to the whole antild repertoire.
Flow microfluorometric analysis of H-2L expression.
Authors: Potter T A , Hansen T H , Habbersett R , Ozato K , Ahmed A
Published: December 2022

The cell surface expression of H-2L, a major transplantation antigen, was compared by flow microfluorometry to the expression of products of H-2K and H-2D loci, using monoclonal antibodies. By this methodology, the ontogeny and tissue distribution of Ld antigens were found to be indistinguishable from those of the K and D antigens. In a reciprocal blocking assay, using fluorescein-labeled test reagents, it was shown that monoclonals anti-H-2.65 and anti-H-2.64 did not inhibit the binding of each other. These results suggest that the alloantigenic determinants H-2.64 and H-2.65 are located at distinct sites on Ld molecules. Quantitative comparisons using the fluorescein-labeled monoclonal reagents indicated that Ld molecules are expressed at 2- to 3-fold lower levels on the cell surface compared with K and D molecules. These findings give new credence to a "3-locus" model for the major histocompatibility complex of man and mouse, where H-2L and HLA-C share several homologies that are unique and distinguish them from the other histocompatibility loci.
A fourth heavy chain variable region subgroup, w, with 2 variants defined by an induced auto-antiserum in the rabbit.
Authors: Roux K H
Published: December 2022

An antiserum recognizing previously undescribed antigenic determinants (designated subgroup w) of the heavy chain VH region was produced by immunizing a rabbit, homozygous for the VH haplotype a1x-y- with IgG from another a1x-y- rabbit, suppressed for a1. The antiserum gives a single fused precipitin band against sera from homozygous rabbits suppressed for the a1, a2, or a3 allotypes. By gel diffusion analysis, the determinant was found to be distinct from previously described a, x, and y allotypes, and was detectable on IgG, F(ab')2 gamma and the gamma-heavy chain. Radioinhibition analysis revealed 2 patterns of inhibition suggesting 2 variants of w that are closely linked to VH allotypes. Variant w34 was detected at very low levels (0.042 to 5.6 micrograms/mg IgG) in normal rabbits expressing the a1x-y-, a1-y33,30, and a3x32y- VH haplotypes. Variant w35 was detectable in a 2x32y33,- rabbits. Sera from homozygous a allotype-suppressed rabbits representing each of the 4 VH haplotypes similarly exhibited either w34 or w35 patterns of inhibition as described above. The level of w34 in a1- and a3-suppressed rabbits exhibited a dramatic 230-fold median increase. In contrast, w35 levels for a2-suppressed rabbits fell within the normal range. In addition to reacting with essentially all normal and suppressed sera, the antiserum reacted with preimmune serum from the same rabbit. Thus, anti-w represents an induced auto-antibody. These observations are further discussed with regard to rabbit and mouse VH populations.
Transplantation in miniature swine. IX. Swine histocompatibility antigens: isolation and purification of papain-solubilized SLA antigens.
Authors: Metzger J J , Gilliland G L , Lunney J K , Osborne B A , Rudikoff S , Sachs D H
Published: December 2022

Histocompatibility antigens have been purified by papain treatment of crude cellular membranes obtained from spleen and mesenteric lymph nodes of individual miniature swine that are homozygous at the major histocompatibility complex. The purification protocol included ultracentrifugation, gel filtration, ion-exchange chromatography, and in some instances preparative isoelectric focusing. This procedure was applied to the preparation of sLA antigens from the 3 haplotypes available in our partially inbred miniature swine herd. The purity of the SLAdd molecules was evidenced by: 1) a single activity peak of approximately 50,000 daltons on gel-permeation chromatography; 2) the antigen elution as a symmetrical peak from ion-exchange resins and from isoelectric focusing columns; 3) the presence of 2 predominant polypeptide chains on SDS-PAGE at apparent m.w. 43,000 and 13,000 daltons; and 4) specific immunoprecipitations by alloantisera and by anti-beta 1 microglobulin antisera. The antigenic activity of SLA products was analyzed by inhibition of complement-mediated cytotoxicity and by removal on anti-beta 2-microglobulin affinity columns. Heavy and light chains were separated by gel-filtration on Sephacryl S-200 in 6 molar guanidine. These SLA products, which have thus been shown to be pure by a variety of criteria, are now readily available for structural analysis and for analysis of biologic activity.
Long-term in vitro culture of murine mast cells. I. Description of a growth factor-dependent culture technique.
Authors: Tertian G , Yung Y P , Guy-Grand D , Moore M A
Published: December 2022

A feeder layer independent long-term in vitro culture system for murine mast cells is described. Concanavalin A-activated murine splenic leukocyte-conditioned medium, prepared under conditions optimal for T cell growth factor production, has been found also to contain a growth-promotion activity for murine mast cells identified by their morphology, characteristic ultrastructure of the granules, positive reactions with toluidine blue and alcian blue, presence of receptors for IgG and IgE, as well as presence of histamine, serotonin, L-Dopa, 5-hydroxytryptophan, and sulfated products within the cytoplasm. After 2 to 3 wk of culture in the presence of the conditioned medium, mast cell lines were established from various sources initially devoid of matured mast cells. Such sources included spleen and bone marrow of athymic nude mice, long-term cultured marrow cells as well as T cell-depleted normal marrow. Cultured mast cells are absolutely dependent upon the conditioned medium-derived growth factor(s) for growth and viability. Death ensues within 24 hr in the absence of the factor(s). Established mast cell lines have been maintained in exponential growth for over 1 yr by passaging in the conditioned medium every 3 to 7 days.
Induction of human immunoglobulin secretion. I. Synergistic effect of B cell mitogen Cowan I plus T cell mitogens or factors.
Authors: Saiki O , Ralph P
Published: December 2022

Synergistic effects of B cell mitogen Staphylococcus bacteria strain Cowan I (Cowan I) plus T cell mitogens in generating immunoglobulin-secreting cells (ISC) from human peripheral blood mononuclear cells (MNC) were investigated. ISC were assayed by reverse plaque-forming cells with protein A-coated red blood cells. Low concentrations of pokeweed mitogen (PWM) plus Cowan I gave superadditive effects on ISC induction, generating 3 to 10 times as many ISC as optimal amounts of either mitogen alone. The mitogens together and separately showed similar kinetics of ISC; synergy was observed at every day tested. Plaque-forming cells of IgM, IgG, and IgA classes all showed strong synergy, together routinely representing 20% of initial MNC. At day 7 of culture, over 80% of non-E-rosetting cells were ISC. Cell donors tested gave these strong responses even if they were low responders to either mitogen alone. Cowan I plus other T cell mitogens, PHA, Con A, and protein A, also provided good signals for B cell activation. Cowan I induced marked proliferation of purified B cells, but T cell-helper signals were required for differentiation to ISC. T cell-helper factor, induced by PWM or PHA, also showed synergistic effects with Cowan I in induction of ISC. Purified B cells did not respond to T cell-helper factor(s) alone to proliferate or differentiate to ISC. These results indicate that optimal ISC induction occurs with a B cell mitogen plus T cell signals acting synergistically.
Characterization of phosphorylcholine- (PC) specific IgE-B cells in CBA/N or (CBA/N x BALB/c)F1 male mice.
Authors: Shigemoto S , Kishimoto T , Yamamura Y
Published: December 2022

Surface immunoglobulin isotype of precursors for PC-specific IgE response in CBA/N or NBF1 male mice was studied. Immunization of CBA/N or NBF1 male mice, which had been lethally irradiated and reconstituted with KLH-primed syngeneic spleen cells, with PC-KLH induced the PC-specific IgE, but not the IgM, response. Depletion of mu-bearing cells from KLH-primed spleen cells completely abolished the induction of the anti-PC IgE response. Depletion of T15 idiotype-bearing cells with anti-T15-coated dishes also abrogated the anti-PC IgE response in NBF1 male mice. Mice that were transferred with delta- or epsilon-bearing cell-depleted cell populations did not show the anti-PC IgE response on day 14, but they showed the anti-PC IgE response on day 21. These results indicated that PC-specific IgE-producing cells in CBA/N mice, which were defective in the anti-PC IgM response, were derived from PC-specific mu-bearing cells, and direct precursors bore mu, delta, and epsilon on their surface.
Increased natural killer cell activity in experimental American trypanosomiasis.
Authors: Hatcher F M , Kuhn R E , Cerrone M C , Burton R C
Published: December 2022

These studies showed that increased natural killer (NK) cell activity develops during experimental American Trypanosomiasis in mice. Increased cytotoxicity against YAC target cells by peritoneal cavity cells (PEC) was detected as early as 1 day into infection in both C3H/He and C57BL/6 mice. Peak activity occurred 2 days into infection, with significant but variable increases detected in the spleen cells (SC) from these mice. The response subsided in PEC and to a lesser extent in SC by 4 days post-infection. Similar responses were not detected in infected C57BL/6 bgJ/bgJ (beige mutant) mice until the fourth day of infection and peaked at significantly lower levels than the peak on day 2 in beige hybrid mice ([C57BL/6 +/+ X C57BL/6 bgJ/bgJ]F1). The effector cell from infected mice was sensitive to pretreatment with anti-NK 1.2 serum plus complement (C), partially sensitive to anti-Thy 1.2 + C, and insensitive to polyvalent anti-mouse immunoglobulin + C. The cytotoxic activity induced in cells from mice infected for 2 days was recovered in subpopulations nonadherent to plastic or Sephadex G-10. Thus, the anti-YAC effector cell found very early in mice infected with T. cruzi possessed many of the characteristics of natural killer (NK) cells found in normal mice. Other experiments demonstrated that injection of heat-killed preparations of blood-form trypomastigotes (BFT) induced increases in NK activity. The presence of augmented NK activity against YAC tumor cells in both resistant and susceptible strains of mice, together with the presentation of the resistance phenotype in beige mutant mice, which had a lower NK response to infection, is not indicative of a direct role for host protection during infection with T. cruzi by NK cells.
How complement kills E. coli. II. The apparent two-hit nature of the lethal event.
Authors: Wright S D , Levine R P
Published: December 2022

We have studied the nature of complement (C) action on red blood cells and E. coli with respect to the number of "hits" required for membrane damage. Our method of analysis involves adding various amounts of purified C7 or C8 to serum preparations immunochemically depleted of C7 or C8, respectively, in order to construct dose-response curves for the action of C's terminal complex. The shape of the dose-response curves reflects the single or multiple-hit nature of C action. Our method confirms that C acts on red cells by a 1-hit mechanism, whether measured by lysis or by the permeation of a small molecule. In contrast, we find with E. coli that C-mediated outer membrane damage, inner membrane damage, and killing all appear to require more than 1 hit. We have also discovered a property of E. coli that displays a nonlethal 1-hit response to C that is particularly useful in the analysis of multiple-hit dose-response curves. Simultaneous measurements of this single-hit phenomenon and the multiple-hit killing of E. coli allow us to make direct comparisons of the amount of C needed for each response. On the basis of the midpoints of the single and multiple-hit curves, C-mediated membrane damage and killing of E. coli appear to be a 2-hit process.
Reciprocal transfer of abnormalities in clonable B lymphocytes and myeloid progenitors between NZB and DBA/2 mice.
Authors: Jyonouchi H , Kincade P W , Good R A , Fernandes G
Published: December 2022

Previous studies in this laboratory revealed that NZB mice have abnormalities of myeloid progenitor populations from an early age such that they are poorly responsive to a particular type of colony-stimulating activity (CSA). In addition, these mice develop abnormally high numbers of B cells that can be cloned in semisolid agar cultures and that are atypical in resisting inhibition by anti-mu antibodies. To investigate whether these abnormalities, like other autoimmune phenomena studied previously, are transferrable with hemopoietic cell grafts, we performed reciprocal bone marrow transplants between NZB and normal DBA/2 mice. Irradiated control mice given syngeneic marrow did not change with respect to any of the parameters that were measured. In contrast, DBA/2 recipients of NZB marrow were indistinguishable from NZB mice in terms of CSA responses and incidences of anti-mu resistant B cells. The proportions but not total numbers of clonable B cells ere elevated in these mice until at least 16 wk after grafting. Conversely, NZB mice given DBA/2 cells had all of the normal characteristics of DBA/2 mice. Transplantation did not cause significant changes in hematocrits, reticulocyte counts, or spleen weights. Therefore, all elements necessary for expression of these lymphoid and nonlymphoid abnormalities of NZB mice are intrinsic to radiosensitive hemopoietic cells.
Deactivation of human neutrophil chemotaxis by chemoattractants: effect on receptors for the chemotactic factor f-Met-Leu-Phe.
Authors: Donabedian H , Gallin J I
Published: December 2022

Normal human peripheral blood PMN were exposed to varying concentrations of partially purified chemotactic complement fragments (C5fr) and a chemotactic peptide N-formyl methionylleucylphenylalanine (f-Met-Leu-Phe). This exposure resulted in a decreased chemotactic response termed deactivation of chemotaxis. Deactivation was found to be nonpreferential for the deactivating stimulus when high concentrations of either f-Met-Leu-Phe (10(-6) M) or C5fr (20 micrograms/ml) were used. When PMN were incubated with lower concentrations of C5fr (10 micrograms/ml), there was preferential deactivation towards C5fr. Similarly, preferential deactivation of chemotaxis was observed when PMN were incubated with 10(-6) M f-Met-Leu-Phe, but this was transient and cells were nonpreferentially deactivated 60 min after the initial exposure to f-Met-Leu-Phe. The availability of receptors for tritiated f-Met-Leu-Phe was examined by Scatchard analyses and measurement of reversible f-Met-Leu-[3H]Phe binding to C5fr and f-Met-Leu-Phe-deactivated PMN. When PMN f-Met-Leu-Phe receptors were studied immediately after exposure to concentrations of C5fr causing either preferential or nonpreferential deactivation, there was increased receptor availability compared with control PMN. In contrast, PMN deactivated with high concentrations of f-Met-Leu-Phe 10(-6) M) had a transient decrease in the number of receptors followed 1 hr later by an increase in the number of receptors. This was similar to the functional correlate of preferential deactivation of chemotaxis immediately after incubation with f-Met-Leu-Phe followed by nonpreferential deactivation in these same PMN. The data indicate that preferential deactivation of chemotaxis may be associated with a preferential decrease (down-regulation) of chemoattractant receptors and that nonpreferential deactivation is associated with an increase in chemoattractant receptors.
Lipid metabolism during mediator release from mast cells: studies of the role of arachidonic acid metabolism in the control of phospholipid metabolism.
Authors: Marquardt D L , Nicolotti R A , Kennerly D A , Sullivan T J
Published: December 2022

Recent studies indicate that both arachidonic acid (AA) metabolism and phospholipid (PL) metabolism are markedly stimulated during the release of mediators from mast cells. The relationship between stimulated AA metabolism and stimulated PL metabolism in isolated rat mast cells was investigated and then correlated with the secretory process. ETYA (5,8,11,14-eicosatetraynoic acid, a known inhibitor of cyclooxygenase and lipoxygenase pathways of AA metabolism) inhibited 32PO4 incorporation into phosphatidic acid (PA), phosphatidylinositol (PI), and phoshodidylcholine (PC) in both unstimulated mast cells and mast cells stimulated by cross-linking of surface IgE molecules. ID50 values for inhibition of mediator release and of basal and stimulated 32PO4 incorporation into PL were 50 to 60 microM ETYA. Indomethacin (1 to 10 microM) and aspirin (10 to 100 microM) had no significant effect on 32PO4 incorporation or on mediator release. AA (10 microM) inhibited PL labeling in resting mast cells and rendered the cells less responsive to secretory signals. Preincubation of the cells with indomethacin (1 microM) blocked both of these AA effects. When AA was added to stimulated mast cells, however, both PL labeling and mediator release were enhanced. Thus, each of the alterations in AA metabolism caused parallel changes in mast cell PL metabolism and in mediator release. Since both basal and stimulated PL metabolism were modified by ETYA and AA, some form of direct regulation of mast cell PL metabolism by AA metabolites seems likely. The close parallelism of effects on mediator release and on PL metabolism suggests that modulation of mast cell function by AA metabolites may be mediated at lest in part by effects on lipid metabolism.
A genetic locus responsible for salmonella susceptibility in BSVS mice is not responsible for the limited T-dependent immune responsiveness of BSVS mice.
Authors: Briles D E , Benjamin W H , Williams C A , Davie J M
Published: December 2022

BSVS mice are known to be highly susceptible to salmonella infection. We have shown that the bulk of the difference in susceptibility between BSVS and salmonella-resistant A/J mice is the result of a genetic difference at a single locus not closely linked to H-2, Igh-C, or Hbb, and not X-linked. We have backcrossed the A/J allele at this locus into BSVS mice for 8 successive generations and have demonstrated that the salmonella resistance afforded by this allele is not the result of a restoration of the generalized poor T-dependent responsiveness of BSVS mice. The salmonella resistance locus we have examined with these 2 strains is probably the same as the Ity locus described by others.
Analysis of Ir gene control of cytotoxic response to hapten-modified self: helper T cells specific for a sulfhydryl hapten can substitute for an anti-TNP-H-2b self helper cell defect.
Authors: Fujiwara H , Levy R B , Shearer G M
Published: December 2022

Helper T cells specific for N-iodoacetyl-N'-(5-sulfonic 1-naphthyl) ethylene diamine (I-AED) were generated in (C56BL/6 X C3H/He)F1 mice by immunization with I-AED-modified syngeneic cells (AED-self). The requirements for activation of hapten-induced helper cells were investigated. The results demonstrated that activation of AED and trinitrophenyl- (TNP) helper cells was strictly hapten specific. In addition, F1 AEd-helpers could be activated efficiently by either I-AED-modified H-2b or H-2k self components to enhance the anti-AED self-CTL responses. This contrasts with the previous findings demonstrating the failure of TNP-H-2b self to activate F1 TNP-helper cells. After AED-helpers were activated, they were capable of augmenting sensitization of cytotoxic T cells (CTL) against TNP-self. These results indicate that although the activation of hapten-reactive helper cells is antigen (hapten)-specific, the subsequent helper activity, as determined by augmentation of CTL responses against another hapten, is antigen nonspecific. Since helper function was antigen nonspecific, F1 AED-helper cells activated by AED-H-2b or AED-H-2k self were tested for their ability to augment the F1 and anti-TNP-H-2b CTL response. The results indicate that the Ir gene defect in the ability of F1 spleen cells to respond to TNP-H-2b self could not be corrected by these helper cells. These results are discussed in the light of Ir gene controlled differences in the activation of AED and TNP-helper cells and possible models for augmenting CTL responses against various antigens in strains that generate marginal helper activity to TNP-self.
Immunoglobulin-bearing cells are a target for the antigen-induced inhibition of pokeweed mitogen-stimulated antibody production.
Authors: Stevens R H
Published: December 2022

In vivo immunization with tetanus toxoid causes the appearance of B cells in the circulation that can produce IgG-antitetanus toxoid antibody in vitro after stimulation by T cells and pokeweed mitogen. Addition of soluble tetanus toxoid to these cultures, however, causes a profound inhibition of the in vitro IgG, but not IgM, antitetanus toxoid antibody production. Treatment of the lymphocytes with 10 microgram of tetanus toxoid for as little as 1 hr at 37 degrees C was sufficient to promote this antigen-specific inhibition. The antigen could be added, however, as late as 3 days after the culture initiation for the inhibition to be observed. Cell separation studies indicated that the inhibitory activity could be found in both the E-rosette positive and E-rosette negative populations. Further studies on the non-E-rosetting cells showed that: 1) inhibition induced in antigen-treated cells could not be transferred to untreated cells; 2) adherent cells were not responsible for the inhibition; 3) treatment of the surface Ig+ cells with antigen was sufficient for the inhibitory effects; and 4) in vivo generated lymphoblastoid cells that produce IgG-antitetanus toxoid antibody in vitro but are surface Ig- were not inhibited by soluble antigen. The results in this study thus indicate that cells in both the E-rosette positive and negative subsets can be responsible for the antigen-induced inhibition of in vitro antibody production observed in the majority of immunized donors.
Lymphocyte recognition of lymph node high endothelium. III. Enhancement by a component of thoracic duct lymph.
Authors: Carey G D , Chin Y H , Woodruff J J
Published: December 2022

The first step in the entry of blood lymphocytes into lymph node is the binding of these cells to venules lined by high endothelium (HEV). This recognition event has been studied using a model system in which rat thoracic duct lymphocytes (TDL) adhere selectively to HEV when overlaid onto lymph node sections. We now report that this binding is increased by a lymph fraction designated adherence enhancing factor (AEF). Lymph depleted of cells and chylomicra was fractionated with (NH4)2SO4. Material that precipitated between 60 and 80% saturation was dissolved in buffer and extracted with chloroform/methanol; the aqueous phase was used as the source of AEF. The factor was nondialyzable, trypsin sensitive, and heat stable (70 degrees C, 30 min). Lymph node sections pretreated with AEF showed a 2- to 3-fold increase in the number of TDL bound to HEV and a 5 to 10-fold increase in the number of HEV with more than 5 adherent lymphocytes. The effect of AEF was specific: 1) No increase in lymphocyte binding to non-HEV areas of the tissue section occurred. 2) The binding of both TDL and spleen lymphoid cells to HEV was enhanced, but thymocyte attachment was not promoted. We conclude that AEF has biologic properties that might enable it to play a physiologic role in lymphocyte recirculation.
Detection and localization of a lectin on Actinomyces viscosus T14V by monoclonal antibodies.
Authors: Cisar J O , Barsumian E L , Curl S H , Vatter A E , Sandberg A L , Siraganian R P
Published: December 2022

A cell-associated lectin activity that mediates lactose-inhibitable adherence of Actinomyces viscosus T14V has been localized to a specific population of fimbriae by the use of monoclonal antibodies. Nine monoclonal antibodies were produced that reacted with only 1 of 2 immunoelectrophoretically distinct fimbrial components on T14V. The fibrillar morphology of this component was revealed by the immunoelectronmicroscopic examination of bacteria incubated with the monoclonal antibodies. The lectin activity associated with these structures was detected when isolated fimbriae were cross-linked with monoclonal antibodies to form immune complexes with agglutination activity for neuraminidase-treated human erythrocytes, a reaction that was inhibited by lactose. Although the 9 monoclonal antibodies differed in their fine specificities, they reacted only with strains of A. viscosus and A. naeslundii that exhibited lactose-inhibitable adherence. These findings indicate that the lectin activity common to these bacteria resides on fimbriae that are antigenically related to those of T14V.
The role of interleukin-2 (IL-2) in the differentiatin of cytotoxic T cells: the effect of monoclonal anti-IL-2 antibody and absorption with IL-2 dependent T cell lines.
Authors: Kern D E , Gillis S , Okada M , Henney C S
Published: December 2022

A variety of approaches, including the use of a monoclonal antibody, have indicated that the lymphokine Interleukin 2 is a necessary and sufficient mediator for initiating cytotoxic T cell differentiation in vitro. Mitogen (Con A, PHA) stimulation of spleen cells and K/D or I region stimulation in primary MLC all led to IL-2 production, as measured by the growth of an IL-2 dependent T cell clone, and to the production of a "helper" factor necessary for cytotoxic T lymphocyte (CTL) differentiation against metabolically inactivated stimulator spleen cells. Several lines of evidence suggested that the IL-2 activity and CTL "helper" activity were due to a single molecular entity. These included the observations that: i) There was a linear relationship (R = 0.99) between IL-2 activity and relative CTL "helper" activity regardless of the preparation used (purified IL-2; MLC, or mitogen-induced supernatants). ii) Absorption of MLC supernatants with T cell "blasts" or with IL-2 dependent T cell lines concomitantly removed both IL-2 and CTL "helper" activities. iii) IL-2 activity purified by isoelectric focusing, or by polyacrylamide gel electrophoresis, displayed CTL "helper" activity. iv) Both CTL "helper" and IL-2 activities were neutralized by a monoclonal antibody directed against IL-2.
Role of accessory cells in B cell activation. IV. Ia+ accessory cells are required for the in vitro generation of thymic independent type 2 antibody responses to polysaccharide antigens.
Authors: Morrissey P J , Boswell H S , Scher I , Singer A
Published: December 2022

It has been previously shown that the in vitro antibody response to TNP-Ficoll requires the presence of adherent accessory cells. In order to determine if this characteristic was unique to TNP-Ficoll or a general feature of the TI-2 antibody responses, responses to the polysaccharide antigens TNP-Levan and TNP-Dextran were studied. Also, it was determined if the functionally relevant accessory cell expresses Ia determinants. Passage of spleen cells over Sephadex G-10 abrogated the response to TNP-Levan and TNP-Dextran as well as to TNP-Ficoll. Addition of adherent accessory cells to the G-10 passed spleen cells reconstituted the response to all 3 antigens. Pretreatment of the adherent accessory cells with a specific anti-Ia serum plus complement abrogated the ability of these cells to provide accessory cell function in the responses to all 3 antigens. Thus, an Ia-positive adherent accessory cell is required for the generation of TI-2 antibody responses to these polysaccharide antigens. This raises the possibility that genetic restrictions may exist between the Ia-positive accessory cell and the lymphocytes involved in the responses to TNP-Ficoll, TNP-Dextran, and TNP-Levan.
A new possible regulatory system for protein phosphorylation in human peripheral lymphocytes. I. Characterization of a calcium-activated, phospholipid-dependent protein kinase.
Authors: Ogawa Y , Takai Y , Kawahara Y , Kimura S , Nishizuka Y
Published: December 2022

A new species of protein kinase (protein kinase C), found in human peripheral lymphocytes, is 20 times more active than adenosine 3',5'-monophosphate-dependent protein kinase (protein kinase A) with histone as phosphate acceptor. This enzyme is normally present as an inactive form in soluble fraction, but attaches to membrane to exhibit full catalytic activity. This activation process is reversible, and absolutely requires Ca++. No other divalent cations can substitute for Ca++. The enzyme is independent of cyclic nucleotides. The active factor associated with membrane is phospholipid. The protein kinase shows a m.w. of 7.7 X 10(4). The pH optimum is 7.5 to 8.5. GTP does not serve as phosphate donor. The enzyme appears to show relatively broad substrate specificity that is distinctly different from that of protein kinase A. Although both protein kinases C and A react with H1 histone, analysis of the N-bromosuccinimide-bisected fragments of this radioactive histone has revealed that protein kinase C greatly favors seryl and threonyl residues of the C-terminal portion, whereas protein kinase A reacts preferentially with seryl residues in the N-terminal portion of this histone. Protein kinase C is protentially multifunctional and may regulate various Ca++-dependent cellular processes. It is noted that protein kinase C may be alternatively activated in an irreversible manner by limited proteolysis with Ca++-dependent neutral protease. The enzyme activated in this way is independent of Ca++ and membrane.
Unresponsiveness in hapten-specific cytotoxic T lymphocytes. I. Characteristics of tolerance induction in adult mice.
Authors: Butler L D , Miller S D , Claman H N
Published: December 2022

These experiments concern the regulation of cytotoxic T lymphocytes (CTL). Here we describe the induction of unresponsiveness in TNP-specific cytotoxic cells generated in vitro. Previous work has shown that i.v. injection of water-soluble TNP (TNBS) prevents in vivo generation of TNP-specific CTL in C3H/HeN mice. In contrast, this report shows that the in vitro generation of TNP-specific CTL from similarly treated C3H/HeN mice is not inhibited. Nevertheless, unresponsiveness in CTL can be induced with TNBS. This unresponsiveness appears to be H-2 linked, since 3 strains of H-2d mice (BALB/c, DBA/2J, and B10.D2) were rendered unresponsive after a single injection of TNBS, whereas 3 strains of H-2k mice (C58/J, C3H/HeN, and CBA/J) were not. This CTL unresponsiveness is antigen specific in both induction and expression. The unresponsiveness is reversed by pretreatment with cyclophosphamide but not by adult thymectomy. Furthermore, the addition of supernatants from Con A-stimulated spleen cells to cultures containing the unresponsive responder cells enhanced the generation of cytotoxic activity. These results suggest that a) the ability to induce TNP-specific CTL unresponsiveness is linked to H-2 loci, b) a regulatory network is involved in the unresponsiveness, which is sensitive to cyclophosphamide but resistant to adult thymectomy, and c) at least 1 level of regulation involves amplifying T cells.
Lymphocyte surface receptors and albumin.
Authors: Sidman C L
Published: December 2022

Albumin was shown to be hidden component of mouse B and T lymphocyte plasma membranes. It was readily radiolabeled from within the plasma membrane by a lipophilic, photoactivated reagent (125I-iodonaphthylazide) but not from the cell exterior by lactoperoxidase-catalyzed iodination; it was not detected by immunofluorescence on intact cells. The function of this cryptic lymphocyte membrane albumin is unknown at present. This cryptic albumin component was discovered during immunoprecipitation experiments using anti-Ig reagents. It is considered of general methodologic significance that many antisera, both native and rigorously Ig-absorbed (both positively and negatively), contained such contaminating activity. The possibility is raised that such contaminating activity may be involved in some reported findings of albumin-size "Ig-like heavy chains" in both the B and T lymphocyte lineages.
Regulatory effect of H2-D region antigens in the graft-vs-host response: F1 hybrid resistance to alloreactive T cells.
Authors: Elkins W L , Quant M M
Published: December 2022

The role of donor and host H2D alleles in the graft-vs-host response was evaluated by measurement of DNA synthetic response of donor lymph node cells in lethally irradiated hosts and by a mortality assay in sublethally irradiated F1 hosts. Comparison of the potency of graded doses of B10.A, B10.A(2R), and (B10.A X B10.A(2R))F1 cells in (B10 vs B10.A)F1 and other hosts expressing foreign KbIb revealed a deficient response of 10(6) B10.A(2R) cells, but only when the host was heterozygous d/b at H2-D. Studies with TI congenic donors failed to implicate T region antigens in this effect. These results are interpreted to suggest that under the conditions of these experiments there is F1 hybrid resistance to alloaggressive T lymphocytes that are homozygous in the Db region.
Spontaneous autoimmune thyroiditis in chickens. I. Effects of bursal reconstitution.
Authors: Polley C R , Bacon L D , Rose N R
Published: December 2022

Chicks from strains genetically susceptible or resistant to spontaneous autoimmune thyroiditis were neonatally treated with cyclophosphamide to induce lymphocytic depletion of the bursa of Fabricius. The subsequent bursal repopulation with B cells from donors matched at the major histocompatibility locus showed B cells to be intricately associated with the pathogenesis of spontaneous autoimmune thyroiditis, although they themselves did not carry the potential for initiation of the disease.
Primary in vitro antibody responses by purified murine B lymphocytes in serum-free defined medium.
Authors: Mosier D E
Published: December 2022

A serum-free medium formulation that allows a primary in vitro antibody response to the nonmitogenic thymic independent antigen TNP-AECM-Ficoll is described. Purified murine B lymphocytes and adherent accessory cells are required for optimal specific responses as well as for optimal B cell survival. The medium is based on Iscove's modification of Dulbecco's modified Eagle's medium and Ham's F-12 nutrient mixture. The serum-replacing components are insulin, transferrin, and progesterone. The survival of unstimulated murine T cells in this Iscove/F12 medium is poor, but B cells survive for relatively long periods of time. This medium should prove useful in defining the molecular requirements for B lymphocyte activation by antigen.
Functions of human monocyte and lymphocyte subsets obtained by countercurrent centrifugal elutriation: differing functional capacities of human monocyte subsets.
Authors: Yasaka T , Mantich N M , Boxer L A , Baehner R L
Published: December 2022

Two monocyte subsets from the peripheral blood of healthy humans were isolated by countercurrent centrifugal elutriation (CCE), and their functional heterogeneity was assessed. Cell volume of the smaller monocyte population was (332 +/- 32 mu 3), compared with the larger monocytes (380 +/- 30 mu 3). Superoxide release by large monocytes exposed to opsonized zymosan was 1.5-fold more active than those of small monocytes. Furthermore, the production of colony-stimulating factor (CSF) and myeloperoxidase activity by the larger monocytes was 2-fold and 1.4-fold greater than that of small monocytes, respectively. Thus, CCE provides a useful means to isolate 2 subsets of monocytes with similar functional properties but quantitatively different activities. This suggests that the 2 subsets may be closely related in their developmental spectrum.
Interferon induces peripheral lymphadenopathy in mice.
Authors: Gresser I , Guy-Grand D , Maury C , Maunoury M T
Published: December 2022

Inoculation (i.p. or i.v.) of partially purified or highly purified (H.P.) mouse interferon (electrophoretically pure in SDS-PAGE gels) into different strains of mice (C3H, BALB/c, Swiss, heterozygote and athymic nu/nu C57BL/6) results in a marked increase in the weight and in the total number of cells in axillary and inguinal lymph nodes. Interferon injected into the hind footpad induced an increase in the weight of the ipsilateral popliteal lymph node compared with the contralateral lymph node. Inoculation of noninterferon proteins (impurities) obtained in the course of purification of this H.P. interferon, mock interferon, or human leukocyte interferon were without effect. Experiments were undertaken to elucidate the mechanism by which interferon induced an increase in the weight (number of cells) of peripheral lymph nodes. There was no evidence that the afflux of transferred chromium-labeled lymphoid cells to lymphoid tissue was increased in interferon-treated recipient mice. There was no evidence that interferon treatment resulted in a stimulation of the multiplication of cells within the peripheral lymph nodes. Since we observed a decrease in the number of lymphocytes in the thoracic duct lymph and in the peripheal blood of interferon-treated mice, we suggest that interferon treatment resulted in an inhibition of egress of lymphocytes from lymphoid tissue. The finding that interferon treatment of mice alters patterns of lymphocyte circulation may be relevant to some of the effects of interferon on the immune system and may explain in part the lymphadenopathy observed in the course of some viral infections.
Identification of a brain theta-positive, secretory cell from hematopoietic tissues.
Authors: Cambier J C , Havran W L , Fernández de Albornoz T , Corley R B
Published: December 2022

We have characterized a population of large, brain-theta antigen-positive, secretory cells that occur in high frequency among murine bone marrow (approximately 6%), spleen (approximately 0.5%), and fetal liver (approximately 5%) cell populations. They do not occur in significant numbers among lymph node, thymus, and peritoneal exudate cells. These cells are Ly1, Ly2, and Thy1.2 negative, adherent to nylon wool, Sephadex G-10, and plastic, but peroxidase and Sudan black negative. They are radiation, cyclophosphamide, and cortisone resistant. These cells secrete a product of 30,000 to 40,000 m.w. of unknown function. Production of antisera specific for this molecule has facilitated identification of the cell by using a reverse hemolytic plaque-forming cell assay. Possible functions of this cell and its product are discussed.