bluuebunny/crossref_metadata_2025 huggingface dataset:
Effects of prostaglandin D2 and theophylline on rat serosal mast cells: discordance between increased cellular levels of cyclic AMP and activation of cyclic AMP-dependent protein kinase.
Authors: Holgate S T , Winslow C M , Lewis R A , Austen K F
Published: December 2022

Previous studies have shown that perturbation of the mast cell IgE-Fc receptor activates adenylate cyclase so as to raise cellular levels of cyclic AMP and to activate cyclic AMP-dependent protein kinase. Theophylline, an inhibitor of cytoplasmic cyclic nucleotide phosphodiesterase, raises cellular cyclic AMP levels, activates Type I and Type II cytoplasmic cyclic AMP-dependent protein kinase isoenzymes, and inhibits immunologic mediator release in a dose-dependent fashion. Since the EC50 values for each of these effects are similar (8 to 9.5 mM), it seems likely that a relationship exists between the activation of cyclic AMP-dependent protein kinase and the inhibition of mediator release. Such inhibition could be due to either to the uncovering of an inhibitory protein by phosphorylation or to the depletion of cyclic AMP-dependent protein kinase holoenzyme, which is essential for productive IgE-Fc receptor-induced activation-secretion coupling. PGD2, which also raises mast cell cyclic AMP levels in a dose-dependent fashion and interacts synergistically with theophylline in this regard, fails to suppress mediator release alone or to add to the inhibitory effect of theophylline. The finding that PGD2 also fails to activate cyclic AMP-dependent protein kinase suggests that the adenylate cyclase stimulated by this agonist is not linked to the mast cell activation-secretion response.
An antigenic determinant of human beta 2-microglobulin masked by the association with HLA heavy chains at the cell surface: analysis using monoclonal antibodies.
Authors: Liabeuf A , le Borgne de Kaouel C , Kourilsky F M , Malissen B , Manuel Y , Sanderson A R
Published: December 2022

Four anti-human beta 2-microglobulin monoclonal antibodies were produced against whole lymphoid cells or against urinary beta 2-microglobulin. Their reactivity was fully inhibited by purified soluble beta 2-microglobulin. The B1.1G6, C23.24.2, and B2.62.2 antibodies bound either to free or to HLA-complexed beta 2m. They recognized the same or very close determinants, since they achieved mutual blocking. In contrast, the C21.48A1 antibody did not bind to the cell surface and did not recognize the same determinant on the purified beta 2-microglobulin molecule as the others. It was able to bind and to form a specific complex with membrane beta 2-microglobulin only, once separated from the HLA heavy chain. These data strongly suggest that the C21.48A1 antigenic determinant might be hidden when the beta 2-microglobulin is complexed with the HLA heavy chains at the cell surface.
Mouse alloantibodies capable of blocking cytotoxic T cell function. III. Studies of the mechanism of blocking of CML by anti-Lyt-2 antibodies.
Authors: Shinohara N , Taniguchi M , Kojima M
Published: December 2022

Attempts were made to analyze the mechanism of action of anti-Lyt-2 antibody in blocking of CML in the absence of complement. Preincubation of killer cells with anti-Lyt-2 antibody followed by washing was found to abrogate the inhibitory effect of the antibody on target cell lysis, whereas an equivalent amount of the antibody present throughout the culture period caused marked inhibition. The failure of pretreatment with anti-Lyt-2 antibody to cause inhibition of CML might be due to rapid turnover of Lyt-2 molecules. Delayed addition of the anti-Lyt-2 antibody to CML cultures decreased the inhibition observed, which indicated that the antibody must be present during early stages of killing in order to exert significant inhibitory effects. The partitioning of CML according to requirements for divalent cations showed that anti-Lyt-2 antibody acts on killer cells during the Mg++-dependent stage of killing. In addition, the antibody was shown to be capable of reversing certain events occurring during this stage. These data are consistent with a role for Lyt-2 molecules during the antigen recognition stage of cell-mediated lysis. However, the exact function of the Lyt-2 molecule remains unknown.
The induction of a unique procoagulant activity in rabbit hepatic macrophages by bacterial lipopolysaccharides.
Authors: Maier R V , Ulevitch R J
Published: December 2022

Disseminated intravascular coagulation (DIC) is a common occurrence during clinical sepsis and can be induced in the experimental host by LPS. Fibrin deposition in the hepatic microcirculation has been observed within 30 min of i.v. injection of LPS. Because mononuclear phagocytes have been shown to produce a PCA after exposure to LPS, we have examined the ability of a homogeneous population of explanted hepatic macrophages to express PCA. Addition of as little as 10 ng/ml of LPS stimulated a 15- to 20-fold increase in PCA over control culture levels within 7 1/2 hr post-treatment. The PCA was found to be membrane-associated, with approximately 90 to 95% of the total PCA present in the cellular lysates, and more than 85% was inhibited by pretreatment of the cells with the diazonium salt of sulfanilic acid, an inhibitor of ecto-enzymes. In contrast to tissue thromboplastin produced by other M phi populations, the H-M phi PCA was found to be markedly sensitive both to heat inactivation at 56 degrees C and to inhibition by 1 mM DFP. Additionally, assays involving both a 1-stage coagulation test as well as an enzyme assay with a Factor Xa-specific substrate (using normal and deficient human plasmas) demonstrated that the H-M phi PCA appears to activate Factor X directly. Unlike tissue thromboplastin, the H-M phi PCA is non-dependent of Factor VII activation. These studies: 1) demonstrate the LPS induces a unique PCA in the H-M phi, and 2) support a role for the H-M phi in the initiation of DIC in endotoxemic shock.
Effect of Leucogenenol on recovery from immunosuppressin by anti-lymphocyte globulins.
Authors: Rice F A , Koo M J
Published: December 2022

Daily treatment of rats with the thymothyroid hormone leucogenenol after their immunosuppression with anti-lymphocyte globulins causes the rats to regain their immunocompetency after 7 days of treatment, in contrast to well over a month, which is required for untreated controls. Immunocompetency was mesured by the ability of the rat to respond to the injection of sheep erythrocytes with the formation of normal concentrations of antibody-forming cells in their lymphoid tissues and normal titers of hemolysins in their serum. These results indicate that leucogenenol increases the rate of development of T cells, as well as B cells, from committed precursors.
Biochemical evidence for trans complementation of structural genes in the expression of I-E antigens in F1 hybrids.
Authors: Plunkett M L , David C S , Freed J H
Published: December 2022

Analysis of the tryptic peptides obtained from the Ia.7-positive, "I-E" antigens produced by 4 F1 combinations derived from strain A.TFR5 has provided biochemical confirmation that I-A/I-E gene complementation can occur in the transconfiguration. The data obtained support the concept that structural genes, as opposed to regulatory genes, are the essential elements involved in this complementation and support the assignment of the I-E subregion of strain A.TFR5 as I-Ek. The data support the following assignments of the genes encoding the E beta chains: B10.A(4R) as E beta k, B10.A(5R) as E beta b, and B10.HTT as E beta s. Finally, data are presented demonstrating that the E beta chain derived from the recombinant D2.GD strain partially differs from the E beta d chain derived from a conventional H-2d haplotype strain, B10.D2.
The Qa-1 alloantigens. I. Identification and molecular weight characterization of glycoproteins controlled by the Qa-1a and Qa-1b alleles.
Authors: Cook R G , Rich R R , Flaherty L
Published: December 2022

Splenocytes from the Qa-Tla congenic strain pairs, A and A-Tlab or B6 and B6-Tlaa, were biosynthetically labeled with 3H-amino acids or cell surface labeled with 125I. Membrane proteins were solubilized with detergent and chromatographed on lentil lectin-Sepharose, and the resulting adherent pools were immunoprecipitated with antisera specific for determinants controlled by the Qa-1a and Qa-1b alleles, Qa-1.1 and Qa-1.2, respectively. Polyacrylamide gel electrophoresis analysis of immunoprecipitates from biosynthetically labeled preparations indicated that both the Qa-1.1 and Qa-1.2 antigens were glycoproteins with a m.w. of approximately 46,000. Qa-1.2 isolated from radioiodinated spleen cells similarly had a m.w. of 46,000. Analysis of anti-Qa-1.1 precipitates from 125I-labeled Qa-1a lysates demonstrated in addition to the 46,000 m.w. component, an electrophoretically heterogeneous protein or series of proteins in the m.w. range of 55,000 to 75,000. The specificity of these reactivities was shown by both antiserum and genetic control immunoprecipitations. These findings indicate that the Qa-1.1 and Qa-1.2 antigens are cell surface glycoproteins that are distinct from the TL antigens, and suggest a further complexity at the Qa-1--Tla locus.
Accessory cell stimulation of T cell proliferation requires active antigen processing, Ia-restricted antigen presentation, and a separate nonspecific 2nd signal.
Authors: Germain R N
Published: December 2022

The roles of Ia+ accessory cells in H-2-restricted stimulation of antigen-specific T cell proliferation were explored in an in vitro model. L-glutamic acid60-L-alanine30-L-tyrosine10-(GAT) primed BALB/c nylon wool-passed T cells were depleted of Ia+ antigen-presenting cells (APC) by treatment with monoclonal anti-Ia antibody plus complement. Such cells failed to respond to soluble GAT, or to soluble GAT in the presence of phorbol myristic acetate (PMA), which is known to stimulate production of, or replace, IL-1 in vitro. Addition of gamma-irradiated syngeneic spleen cells reconstituted the response to soluble GAT, but addition of ultraviolet (UV) light-irradiated spleen cells did not, even in the presence of PMA. Preincubation of cells with GAT for 24 hr, followed by washing, then gamma irradiation, generated a cell population able to stimulate GAT-primed T cells to proliferate. The same pulsed cells exposed to UV irradiation failed to stimulate T cell responses unless PMA was added to the cultures. The relevant cells in this UV-irradiated population are Ia+. It is concluded that a finite period of time for interaction of metabolically intact APC with antigen is required before creation of an appropriate (Ia + antigen) signal recognized by the T cell. In addition to such Ia-restricted antigen presentation, however, a 2nd nonspecific signal, again requiring metabolically active APC for elaboration, is necessary for detectable T cell activation. These studies thus define 3 separable activities of APC during the process of H-2 restricted T cell activation.
Immunoglobulin glycopeptides from an IgG2b-producing mouse myeloma cell line and from variant cell lines.
Authors: Weitzman S , Portmore J
Published: December 2022

A mouse myeloma cell line, 45.6.3, produces an IgG2b immunoglobulin (Ig) with 2 carbohydrate attachment sites on the heavy chains. One site is in the CH2 domain and the other in the VH region. The oligosaccharides at each site have different structures. The ratio of radioactive glucosamine incorporated in the VH compared with the CH2 oligosaccharide is approximately 1 to 3. In an attempt to understand this observation further, variant cell lines derived from 45.6.3 were isolated and their Ig were characterized. A ricin-resistant line, R4R1.5, has the same 2 attachment sites as the wild type, but the ratio of radioactive glucosamine in VH compared with CH2 was 1:1 and not 1:3 as in the wild type. This alteration is most probably due to cellular factors, since the Ig protein is unchanged. The M3.11 cell line produces an Ig with a polypeptide deletion involving the CH3 domain. In this Ig, a 3rd carbohydrate attachment site can be demonstrated. The percentage of radioactivity glucosamine in the CH2 domain compared with the total Ig is about 25% instead of 75% as in the wild type. These results suggest that the extent of glycosylation of different sites on Ig can be affected by both cellular factors and structural changes in the Ig protein.
Stimulation with autologous lymphoblastoid cell lines: lysis of Epstein-Barr virus-positive and -negative cell lines by two phenotypically distinguishable effector cell populations.
Authors: Zarling J M , Dierckins M S , Sevenich E A , Clouse K A
Published: December 2022

Human lymphocytes, stimulated in vitro for 6 days with x-irradiated or glutaraldehyde-treated autologous Epstein-Barr (EB) virus-transformed lymphoblastoid cell lines (LCL), are cytotoxic for autologous and allogeneic EB+ LCLs as well as for several EB- cell lines that are also susceptible to lysis by interferon-activated natural killer (NK) cells. To determine whether the apparent nonspecific lysis mediated by LCL-stimulated cells is due to a mixture of effector cells directed against different target cells, advantage was taken of our recent finding that monoclonal antibody OKT8 reacts with human cytotoxic T lymphocytes but not with NK cells or NK-like cells generated in mixed leukocyte cultures. The depletion of OKT8+ cells from LCL-stimulated cultures by treatment with OKT8 and complement abolished or markedly depleted cytotoxicity against all EB+ target cells tested, whereas cytotoxicity against EB-, NK-sensitive cell lines including K562, MOLT-4 and HSB-2 was not or only minimally reduced. These results indicate that stimulation with autologous LCL results in the generation of OKT8+ cytotoxic T lymphocytes that lyse EB virus-transformed LCL and OKT8- NK-like cells that lyse EB-, NK-sensitive cells.
Effect of murine tumors upon delayed hypersensitivity to dinitrochlorobenzene. III. In vivo activity of the nonspecific suppressor cell.
Authors: Jessup J M , Macek C M , Kahan B D , Pellis N R
Published: December 2022

Tumor-induced immunosuppression was investigated in an in vivo model of delayed hypersensitivity (DH) to the chemical sensitizer, dinitrochlorobenzene (DNCB). DH to DNCB as measured in a footpad assay was decreased in C3H/HeJ mice bearing MCA-F, a 3-methylcholanthrene-induced syngeneic fibrosarcoma. Suppressor cells from the spleens of tumor-bearing mice inhibited the induction of DH to DNCB in otherwise normal syngeneic C3H/HeJ recipients. Ten million spleen cells (SpC) harvested from mice bearing MCA-F for 10 days and adoptively transferred to tumor-free mice at the time of sensitization with DNCB suppressed the response to the sensitizer. The suppressor cells were macrophages, since they were adherent to plastic, removed by treatment with a magnet after phagocytosis of carbonyl iron, resistant to exposure to gamma radiation and to treatment with anti-Thy 1.2 serum and complement. Further, the nonspecific suppressor cells were activated by progressive tumor growth rather than by induction of tumor-specific immunity using irradiated tumor cells. Titration studies revealed that suppression of DH occurred with the transfer of as few as 10(6) SpC. Thus, nonspecific suppressor cells are effective at inhibiting in vivo DH to DNCB and suggest that nonspecific suppression in the intact host occurs through mechanisms different from those involved in suppression in vitro.
Natural killing (NK) and antibody-dependent cellular cytotoxicity (ADCC) in specific pathogen-free (SPF) miniature swine and germfree piglets. III. Two distinct effector cells for NK and ADCC.
Authors: Huh N D , Kim Y B , Amos D B
Published: December 2022

Effect of "anti-NK" sera on NK and ADCC activities was examined to study further the relationship between the effector cells for NK and ADCC in specific pathogen-free miniature swine. An appropriate dilution of "anti-NK" sera that had no effect on ADCC activity not only inhibited NK activity but also eliminated NK cells in the presence of C which supports the presence of NK cells distinct from K cells for ADCC.
Ia specificities on parental and hybrid cells of an I-A mutant mouse strain.
Authors: Hansen T H , Walsh W D , Ozato K , Arn J S , Sachs D H
Published: December 2022

Splenic B cells and B cell blasts from the I-A mutant mouse strain B6.C-H-2bm12 were tested by serology with a series of new monoclonal anti-Iab antibodies. Four out of 5 of those monoclonal antibody-defined specificities that are determined by wild-type I-Ab antigens were undetectable on B6.C-H-2bm12 cells. Specificities both present and absent on mutant cells appear to be determinants on the same wild-type molecule, as indicated by sequential precipitation experiments with soluble H-2b antigens. The lack of expression of certain Ia specificities on mutant cells was found not to be the result of disparate control by the Xid gene, which was previously shown to control the expression of Ia.W39, another specificity absent in B6.C-H-2bm12 mice. Serologic testing of Ia specificities on cells and blasts from F1-hybrid mice suggested that the Iabm12 antigens are codominantly expressed, indicating a failure to detect trans regulation or complementation of the mutant phenotype. Another monoclonal antibody-defined Ia specificity dependent on the expression of the E beta polypeptide was normally expressed in B6.C-H-2bm12 mice. These data thus suggest that the lesion of these mutant mice occurred in the A alpha and/or A beta structural gene, resulting in the loss of several Ia specificities.
Membrane expression of Fc-receptors in cultured leukemic cell lines. I. Induction of Fc-receptor in undifferentiated types of cells after passive modulation of lipid viscosity.
Authors: Kyoizumi S , Kannagi R , Masuda T
Published: December 2022

The role of lipid environment of plasma membrane for the expression of Fc receptor (gamma) (FcR gamma) were studied by physicochemical modification of the membranes of myeloid leukemia cell line cells, M1. Sterol binding polyene antibiotics such as amphotericine B or filipin inhibit the expression of FcR gamma during the differentiation of M1- cells by the CM obtained from the culture of mouse embryonic fibroblasts, suggesting the possible involvement of cholesterol molecules in the induction mechanism of FcR gamma. Low-temperature incubation at either 20 degrees C or 4 degrees C results in the increment of their membrane microviscosity, when tested by the fluorescence depolarization method, in company with the appearance of FcR gamma on approximately 60% of M1 cells when assayed by the EA-rosetting method. FcR gamma is also induced on approximately 40% of M1- cells by incubating cells with cholesterol-phosphatidyl-choline liposomes with the concomitant increase of their membrane microviscosity. Nevertheless, no phagocytosis of EA is observed in these cells treated with such physicochemical procedures. These findings imply that FcR gamma is embedded in the membrane of M1- cells and that physicochemical modification of the membrane lipid bilayers can induce the FcR gamma independent on their functional differentiation marked by the expression of phagocytic activity.
Inhibition of T cell-mediated functions by MVM(i), a parvovirus closely related to minute virus of mice.
Authors: Engers H D , Louis J A , Zubler R H , Hirt B
Published: December 2022

A purified preparation of MVM(i), a murine parvovirus closely related to minute virus of mice (MVM), was found to inhibit various functions mediated by murine T cells in vitro. Addition of MVM(i) virus to secondary allogeneic mixed leukocyte cultures resulted in the inhibition of both lymphocyte proliferation (3H-thymidine incorporation) and the generation of cytolytic T lymphocyte activity but not interferon production. MVM(i) virus also inhibited the growth and cytolytic activity of several cloned, long-term Lyt-2+ cytolytic T cell lines. Furthermore, the antigen-induced proliferative responses of parasite- (Leishmania) specific Lyt-1+ T cells in vitro was abrogated by the addition of MVM(i) virus to the culture. Finally, the suppression of an in vitro antibody response to SRBC by MVM(i) virus was the result of the inhibition of T helper cells required for the B cell response. These suppressive effects were specific for MVM(i); parallel studies in which the prototype MVM parvovirus was used showed no significant inhibition in the various systems tested.
Accessory cells in the in vitro generation of type C virus-specific T killer lymphocytes. II. Demonstration of a T helper cell in the spleen of in vivo primed mice.
Authors: Gomard E , Wybier-Franqui J , Simmler M C , McKenzie I F , Levy J P
Published: December 2022

The existence of a helper T cell cooperating with cytolytic T lymphocytes (CTL) in cell-mediated anti-tumor responses specific for the virus-induced FMR antigens can be demonstrated by using unprimed thymocytes as CTL precursors and in vivo primed irradiated spleen cells as helper. The helper T cells express Thy-1.2 and Lyt-1.2 antigens at their surface, but not Lyt-2.2. The helper function required the presence of macrophages to be detected, is antigen specific, and appears unusually radiosensitive compared with previously described helper T cell function.
T cell-mediated cytotoxicity induced by Listeria monocytogenes. I. Activation of Listeria antigen-responsive T cells.
Authors: Woan M C , McGregor D D
Published: December 2022

Soon after rats are infected with Listeria monocytogenes (LM), Listeria antigen- (LMA) responsive lymphocytes are delivered to the animal's thoracic duct. These LM-responsive lymphocytes can be restimulated in vitro by LMA to generate cells that have a potent cytolytic capability. The activation of LMA responsive lymphocytes is immunologically specific and dependent upon the activity of histocompatible accessory cells. Neither cell-free LMA nor LMA-pulsed allogeneic accessory cells can promote a significant cytotoxic response by negatively selected responder lymphocytes. LM-dependent cytolytic lymphocytes differ from natural killer (NK) cells inasmuch as their activation is not facilitated by interferon (IF). Likewise, supernatants from cultures containing specifically sensitized thymus-derived (T) lymphocytes and histocompatible LMA-pulsed accessory cells fail to augment (day 2 and day 3 supernatants actually inhibit) the activation process. The results imply that the successful activation of LM dependent cytolytic lymphocytes requires the cooperative interplay of responder T cells and specific-antigen-pulsed accessory cells.
Suppression of parasite antigen-specific lymphoid blastogenesis in African trypanosomiasis.
Authors: Charoenvit Y , Campbell G H , Tokuda S
Published: December 2022

Inbred C57BL/6J mice were infected with either Trypanosoma rhodesiense organisms of Walter Reed Army Trypanozoon antigenic type 3 or 5 (WRATat 3 or WRATat 5) or were immunized with soluble trypanosomal antigens. Spleen cells obtained from immunized hosts undergo blastogenesis, measured by thymidine incorporation, when exposed to trypanosomal antigens in vitro. Spleens obtained from mice infected with T. rhodesiense organisms do not respond or respond only minimally to trypanosomal antigens in vitro. Spleen cells of infected mice suppress the trypanosomal antigen-induced proliferative response of spleen cells from immunized mice in co-culture experiments. The suppressive activity was found in both the plastic adherent and plastic nonadherent spleen cell populations. The in vitro responses of normal spleen cells to LPS and Con A were also suppressed by spleen cells obtained from infected mice.
Immune responsiveness of SM/J mice: hyper NK cell activity mediated by NK 1+ Qa 5- cells.
Authors: Clark E A , Engel D , Windsor N T
Published: December 2022

Compared with other inbred strains, SM/J mice have both abnormally high responses to B cell mitogens and hyper NK cell and K cell activity. This hyper NK cell activity is evident even in older SM/J animals, and a high proportion of SM/J nylon-nonadherent lymphocytes that bind target cells also have lytic activity. The SM/J NK cells are NK 1+, Qa 5-, but H-2v identical B10.SM congenic mice have normal NK activity and NK 1+, Qa 5+ NK cells. Together these results suggest the elevation of NK cell activity in SM/J mice is due to chronic activation or maintenance of a Qa 5- NK cell subset. The genetic control of B cell responsiveness and NK cell activity was examined in (B6 x SM)F1 x B6 backcross progeny and (A x SM)F2 intercross mice. No correlation between NK levels and mitogen responsiveness was detected. NK levels in SM/J mice appear to be under polygenic, non-H-2 gene control.
Demonstration of a circulating suppressor factor of thymocyte proliferation during endotoxin fever in humans.
Authors: Dinarello C A , Rosenwasser L J , Wolff S M
Published: December 2022

Sub-pyrogenic levels of human leukocytic pyrogen (LP) have been shown to enhance phytohemagglutinin-induced murine thymocyte proliferation. It was concluded that LP is similar to lymphocyte-activating factor (LAF). Since endotoxin stimulates the production of LP and LAF, we attempted to employ in vitro thymocyte proliferation to detect circulating LP/LAF in 12 normal human subjects during experimental endotoxin fever. Sera obtained before and during fever were first mixed with an immunoadsorbent that binds human LP/LAF, and then the dissociated material was added to thymocyte cultures. Material derived from sera obtained during the maximum fever (3 to 4 hr after endotoxin) was markedly suppressive for thymocyte proliferation in vitro. The appearance of this suppressive effect correlated with the profound lymphopenia observed in the subjects. This suppressor factor(s) was nondialyzable and was destroyed at 70 degrees C, and its suppressive effects inhibited the lymphocyte-activating property of LP/LAF. In addition, the suppression of PHA responses did not appear to be modulated by prostaglandin synthesis. The results demonstrate that a factor circulates during endotoxin fever in humans that suppresses in vitro thymocyte proliferation.
Enhancement of T. lymphocyte functions by Fc fragments of immunoglobulin. II. Augmentation of the cell-mediated lympholysis response occurs through an Lyt-1+2- helper T cell.
Authors: Morgan E L , Thoman M L , Weigle W O
Published: December 2022

Fc fragments derived from papain digestion of human IgG have the capacity to augment specific T cell-mediated cytolytic responses. The addition of Fc fragments to cultures containing nylon wool-purified effector and irradiated stimulator cells results in a significant enhancement of the cytotoxic response. Fc fragment-mediated enhancement is restricted to those responses in which the allogeneic differences between effector and stimulator populations encompass the H-2 I region. Significant enhancement of the CML response occurs when differences at H-2 K + I + D (B10 alpha B10.BR) and H-2 I (A.TH alpha A.TL) regions are employed. In contrast strain combinations in which only the H-2 D region (B10.A alpha B10.A(2R)) difference occurs no enhancement is seen. The cellular site of Fc adjuvant action in the CML response is the Lyt-1+2- helper T cell. This was concluded from the finding that Lyt-1+2- helper cells had to be present to obtain Fc-augmented CML responses. The substitution of Interleukin 2 for Lyt-1+2- helper T cells maintained the cytotoxic response, but these responses were not susceptible to the enhancing properties of Fc fragments. Moreover, to achieve enhanced cytotoxic response Fc had to interact only with the cell population.
Ir gene control of the cytotoxic T lymphocyte response to Sendai virus: H-2k mice are low responders to Sendai.
Authors: Shapiro M E , Burakoff S J , Benacerraf B , Finberg R W
Published: December 2022

H-2k mice generate a secondary in vitro cytotoxic T lymphocyte response to Sendai virus 20- to 100-fold weaker than those of other haplotypes tested (H-2b,d,q,s). This immune response defect maps to both H-2K and H-2D. H-2k x H-2d F1 mice (responder x nonresponder) only lyse targets that have the d allele at H-2K and/or H-2D. H-2k targets are equally lysable with anti-Sendai antibody. Furthermore, H-2k mice demonstrate normal antibody and T cell proliferation responses to Sendai virus. The Ir gene defect therefore appears to be limited to the generation of the cytotoxic T lymphocytes.
IgA-specific helper activity of T alpha cells in human peripheral blood.
Authors: Endoh M , Sakai H , Nomoto Y , Tomino Y , Kaneshige H
Published: December 2022

A subpopulation of human T lymphocytes that express receptors of immunoglobulin A (T alpha cells) was separated by a FACS using FITC-conjugated human IgA myeloma protein in order to determine whether they have helper activity for in vitro immunoglobulin synthesis in comparison with a T alpha cell population separated by rosette formation using PTN-ORBC conjugated with MOPC-315 mouse IgA myeloma protein. FACS-separated T alpha cells have IgA specific helper activity, whereas rosette-separated T alpha cells have polyclonal helper activity for in vitro immunoglobulin synthesis in PWM-stimulated B cells. The polyclonal helper activity observed in rosette-separated T alpha cells might be due to contamination of non-T alpha cells in the T alpha cell fraction. It is concluded that purified T alpha cells in normal human peripheral blood show IgA-specific helper activity in vitro.
HLA A, B, C antigens are expressed on nonvillous trophoblast of the early human placenta.
Authors: Sunderland C A , Redman C W , Stirrat G M
Published: December 2022

Samples were taken from the human chorionic sac at 6 wk postconception and were stained with monoclonal antibodies to trophoblast and to HLA antigens by means of an indirect immunoperoxidase technique. Antibody to HLA A, B, C antigens did not stain chorionic villous trophoblast in accordance with previous studies, but did stain nonvillous trophoblast of the cytotrophoblast cell columns and cytotrophoblastic shell. Antibody to HLA DR antigens showed no staining of trophoblast. This is the first demonstration of HLA antigens on a human fetal tissue in direct maternal contact. We have discussed the immunologic implications of this finding.
The antigen-induced selective recruitment of specific T lymphocytes to the lung.
Authors: Lipscomb M F , Lyons C R , O'Hara R M , Stein-Streilein J
Published: December 2022

The purpose of the present studies was to investigate the mechanisms by which specific T lymphocytes accumulate in the lung. After the intratracheal (IT) inoculation of influenza virus into guinea pigs, the detection of specific T lymphocytes in the lung coincided with the development of immunity in both hilar nodes and systemic lymphoid tissue. Animals immunized in the footpads with virus failed to develop immune responses in the lung unless rechallenged IT with immunogen. In adoptive transfer experiments, IT inoculation of influenza into nonimmune guinea pigs, followed immediately by the i.v. injection of a mixture of 3H-thymidine-labeled syngeneic T lymphocytes specific for influenza virus and 14C-thymidine-labeled syngeneic T lymphocytes specific for an irrelevant antigen resulted in the selective accumulation of the virus-specific T lymphocytes in the lung. Taken together, these studies indicate that the selective recruitment by antigen of circulating immune cells is one of the mechanisms by which specific T cells accumulate in the lung.
Biochemical characterization of regulatory factors derived from T cell hybridomas and spleen cells. I. Separation of T cell growth factor and T cell replacing factor from granulocyte-macrophage colony-stimulating factor.
Authors: Clark-Lewis I , Schrader J W
Published: December 2022

We describe the molecular characteristics of T cell growth factor (TCGF), T cell replacing factor (TRF), and granulocyte-macrophage colony-stimulating factor (GM-CSF) produced by a T cell hybridoma after stimulation with concanavalin A (Con A). All three activities could be separated from Con A itself by ammonium sulfate precipitation. The TRF and TCGF activities had a m.w. of 35,000 to 40,000 on gel filtration in phosphate-buffered saline (PBS). Their m.w. were about 30,000 under dissociating conditions in guanidine hydrochloride and about 35,000 to 40,000 under disulfide reducing conditions, suggesting the molecule(s) lacked noncovalent or disulfide-linked subunit structure. GM-CSF had a m.w. of 25,000 to 30,000 by gel filtration in PBS and about 23,000 in guanidine hydrochloride. TRF and TCGF on the one hand and GM-CSF on the other could be distinguished by the criteria of m.w., relative heat sensitivity, and hydrophobic chromatography. TCGF could not be separated from TRF by any of these methods. In terms of all the above properties, factor derived from the T cell hybridoma and spleen cells appeared identical.
Biochemical characterization of regulatory factors derived from T cell hybridomas and spleen cells. II. Evidence for glycosylation of T cell growth factor, T cell replacing factor, and granulocyte-macrophage colony-stimulating factor.
Authors: Clark-Lewis I , Schrader J W
Published: December 2022

Isoelectric focusing demonstrated that T cell growth factor (TCGF), T cell replacing factor (TRF), and granulocyte-macrophage colony-stimulating factor (GM-CSF) derived from concanavalin A-stimulated T cell hybridomas and spleen cells are heterogeneous with respect to charge. The spleen cell-derived TCGF and TRF activities focused with isoelectric points (pI) between 3.5 and 6.5 whereas the range for GM-CSF activity was broader (pI, 3.5 to 8.0). The T cell hybridoma-derived activities were slightly more acidic. Neuraminidase treatment of both hybridoma 123 and spleen cell-derived material resulted in a major peak of each activity (TRF/TCGF pI, 4.9; GM-CSF pI, 4.7). Neuraminidase treatment of hybridoma T6-derived material resulted in peaks of TRF and TCGF around 6.0 as well as one around 5.0, suggesting that this charge heterogeneity was due to causes other than variations in the level of sialic acid on the relevant molecules. Tunicamycin-treated spleen cells or hybridoma 123 cells released biologically active TCGF, TRF, and GM-CSF. Each of these three activities from tunicamycin-treated spleen cells focused with pI around 5.0. A major fraction of TRF, TCGF, and GM-CSF activities bound to wheat-germ agglutinin. GM-CSF also bound to concanavalin A and lentil lectin. These results suggest that the molecules responsible for TCGF, TRF, and GM-SCF activities are glycosylated and that the observed heterogeneity in charge and lectin-binding characteristics is due in part to variable glycosylation. Glycosylation was not critical for any of the three biologic activities. No conclusive separation of TRF and TCGF activities was observed in these experiments although GM-CSF differed from TRF and TCGF in that it bound to Concanavalin A.
Human T cell subpopulations defined by a monoclonal antibody. II. Evidence for cell cooperation in the response to alloantigens and generation of cytolytic cells.
Authors: Moretta A , Corte G , Mingari M C , Moretta L
Published: December 2022

It has been shown previously that the 5/9 monoclonal antibody defines a small T cell subpopulation in human peripheral blood that includes all the cells responsible for proliferation to tetanus toxoid and to alloantigens as well as the helper cells for B cell differentiation. In the present study, human peripheral blood T cells were fractionated according to their reactivity with the 5/9 monoclonal antibody and stimulated in mixed lymphocyte culture (MLC). In spite of a strong proliferative response in MLC, 5/9+ cells generated no cytolytic activity against PHA-activated lymphocytes bearing the stimulating alloantigens (CTL activity) or against the K562 human cell line (NK activity). The precursors of these cytolytic effector cells were present in the 5/9- fraction. However, 5/9+ cells or soluble factors derived from 5/9+ cells were needed to induce 5/9- cells to respond in MLC and develop cytolytic activity. Both 5/9+ and 5/9- cell populations, upon MLC stimulation, were able to lyse L1210 mouse lymphoma cells in the presence of specific antibodies (ADCC).
Production of monoclonal antibodies against MM antigen: the serologic identification of MM antigen with Ly-6.2 alloantigen.
Authors: Seto M , Takahashi T , Tanimoto M , Nishizuka Y
Published: December 2022

Monoclonal antibodies against MM46, an ascitic mouse mammary tumor of C3H/H2, were produced by fusing mouse myeloma cell line NS-1 with spleen cells from a (BALB/c X C3H/HeN)F1 mice hyperimmunized with MM46, an MM antigen-positive tumor. Eight antibodies showed cytotoxicity against MM46, but not against MM48, an MM antigen-negative ascitic mammary tumor, and one hybridoma produced an agglutinating antibody. One of the cytotoxic monoclonal antibodies, 3-3-C, was selected, and the strain distribution and the tissue distribution of MM antigen were studied. The results demonstrated that MM antigen had a strain distribution identical to Ly-6.2 antigen, and a similar tissue distribution. Therefore, the characterization of MM antigen and Ly-6.2 antigen was investigated. Ly-6.2 antibody was shown to be cytotoxic for MM46, but not for MM48, in accordance with 3-3-C. Genetic segregation analysis of MM and Ly-6.2 antigens in 33 backcross mice demonstrated complete concordance between these two antigens. In addition, MM antigen phenotype of an Ly-6.2 congenic strain, C3H.B6-Ly-6b, was studied, and it was found to be positive in contrast to C3H/HeN. Furthermore, cross-absorption studies revealed that both MM46 cells and C3H.B6-Ly-6b lymph node cells could absorb cytotoxic activities of 3-3-C and monoclonal anti-Ly-6.2 antibody. The results so far obtained suggested strongly that these two loci controlling expression of MM and Ly-6.2 antigens were identical or very closely linked.
Idiotypes of Lewis rat antibodies to encephalitogenic peptides of guinea pig myelin basic protein: in vitro and in vivo studies.
Authors: Fritz R B , Desjardins A E
Published: December 2022

Lewis rat antibodies raised by immunization with encephalitogenic peptide 68-88 guinea pig myelin basic protein were purified by affinity chromatography and used to immunize rabbits. After exhaustive absorption of the rabbit antisera to remove anti-rat immunoglobulin activity, the antisera retained activity against the immunogen, shown by the ability to block reaction of radioiodinated peptide with the active site of the rat anti-peptide antibodies. Intrastrain idiotypic cross-reactivity was assessed by testing the rabbit antisera against a panel of Lewis anti-peptide antibodies. Each anti-idiotypic antiserum displayed a unique pattern of reactivity with the panel. Similar tests in which a panel of anti-peptide antibodies raised in F344 rats was used demonstrated the presence of interstrain cross-reactive idiotopes. When seven rabbit anti-idiotypic antisera were tested by pretreatment of rats before challenge with encephalitogen for effect in vivo, five were without effect. Of the remaining two, one caused a slight suppression of disease; the other enhanced disease compared to control animals.
Synovial fluid lymphocytes differ from peripheral blood lymphocytes in patients with rheumatoid arthritis.
Authors: Fox R I , Fong S , Sabharwal N , Carstens S A , Kung P C , Vaughan J H
Published: December 2022

Peripheral blood (PBL) and synovial fluid lymphocytes (SFL) from 12 patients with rheumatoid arthritis (RA) were analyzed using monoclonal antibodies that detect specific T cell surface antigens (OKT3, OKT4, OKT8, SC1) and antigens associated with lymphocyte activation (anti-Ia, OKT10, B3/25). RA patients' PBL contained 1010 +/- 74 OKT4+ and 401 +/- 62 OKT8+ cells/mm3 (ratio OKT4+/OKT8+ = 2.4 +/- 0.3). In contrast, SFL from these patients exhibited a significantly different proportion of T cell subsets (ratio OKT4+/OKT8+ = 1.1 +/- 0.5) (p less than 0.0001) with 630 +/- 180 OKT4+ and 595 +/- 225 OKT8+ cells/mm3. Synovial fluid contained significantly more activated T cells based on the presence of Ia-positive T cells (19 +/- 5%) and reactivity with antibody OKT10 (49 +/- 7%) compared with RA-PBL (8 +/- 3% Ia-positive T cells and 13 +/- 6% OKT10+). Compared with RA-PBL, normal PBL contained an elevated number of OKT8+ cells (610 +/- 48/mm3), a similar number of OKT4+ cells (1040 +/- 86/mm3), and a lower percentage of activated lymphocytes (3 +/- 2% Ia-positive T cells and 10 +/- 8% OKT10+ cells). SFL, RA-PBL, and normal PBL all showed less than 3% cells reactive with antibody B3/25 (anti-transferrin receptor antibody), a marker found on in vitro activated T cells. These findings demonstrate that the lymphocytes at the site of inflammation differ significantly from the lymphocytes present in the peripheral blood of the same patients. RA patients had a small but significant decrease in the number of OKT8+ cells/mm3 compared with normal PBL (p less than 0.01), suggesting that this lymphocyte subset may home to synovial tissues, where it becomes activated.
Kinetics of the anti-recipient cytotoxic cell response of mice with minor histocompatibility antigen graft-vs-host disease.
Authors: Hamilton B L , Parkman R
Published: December 2022

The kinetics of the anti-recipient cytotoxic cell response of spleen cells from mice undergoing graft-vs-host disease (GVHD) induced to minor histocompatibility antigens were studied. Two population of cytotoxic cells were identified. Cytotoxic T lymphocytes (CTL) were present in recipient spleens 2 and 3 wk after transplantation but disappeared from the spleens before the onset of clinical disease. Cytotoxic T lymphocyte precursors (CTLp) were first detected in recipient spleens 2 wk after transplantation and were present during clinical disease. CTL may function as effectors in GVHD induced to minor histocompatibility antigens.
The identification and mapping of a second class I locus in the major histocompatibility complex of the rat.
Authors: Kunz H W , Gill T J , Misra D N
Published: December 2022

A combined genetic, serologic, and biochemical study has demonstrated the existence of a second locus, RT1.E, that codes for class I antigens in the major histocompatibility complex (MHC) of the rat, and has mapped it relative to the other loci in the MHC. The evidence was obtained with the use of two laboratory-derived recombinants, r10 and r11, and the congenic strains DA.1I and BN.1I. Antibodies raised against the i haplotype (RT1.AnBa) in strain combinations that were matched for their RT1.A and RT1.B loci and in the BY1 and anti-r11 strain combination, in which the RT1.A and RT1.B loci and many of the non-MHC genes were matched, reacted with red blood cells and lymphocytes carrying the u haplotype. Immunoprecipitation showed that these antisera reacted specifically with a component of red blood cell membranes having a m.w. of 45,000. Mapping studies place the genes in the rat MHC in the order RT1.ABE; hence, the organization of the MHC in the rat appears to be similar to that of the mouse.
Heterogeneity of human T4+ inducer T cells defined by a monoclonal antibody that delineates two functional subpopulations.
Authors: Reinherz E L , Morimoto C , Fitzgerald K A , Hussey R E , Daley J F , Schlossman S F
Published: December 2022

A monoclonal antibody termed anti-T4 that detected approximately 60% of peripheral blood T lymphocytes was shown to define the human inducer population. In the present study, we characterized three additional monoclonal antibodies, anti-T4A, anti-T4B, and anti-TQ1, that were reactive with a similar percentage of T lymphocytes. Anti-T4A, anti-T4B, and anti-T4 delineated identical cell populations, while those defined by anti-TQ1 differed in several respects: 1) Anti-TQ1 stained a minority (less than 7%) of thymocytes, whereas the other antibodies stained a majority (80%); 2) Anti-TQ1 reacted with 70 to 85% of T4+ lymphocytes, but also stained 50% of T cells within the T4- (T8+) cytotoxic/suppressor subset; 3) The antigen defined by anti-TQ1 was not restricted in its expression to T cells; it defined a fraction of normal B and null lymphocytes as well as non-T cell lines. In vitro studies indicated that the subpopulations of T4+ T lymphocytes delineated by anti-TQ1 were functionally distinct. Although T4+TQ1+ and T4+TQ1- T cells proliferated in an equal fashion to soluble antigen and alloantigen, only the T4+TQ1+ subset was responsible for maximal proliferation in autologous MLR. This T4+TQ1+ subset contained a population of lymphocytes reactive with the previously defined JRA autoantibody. In contrast, the T4+TQ1-, but not the T4+TQ1+, subset provided the majority of T cell help for B cell immunoglobulin production in a pokeweed-driven system. We conclude that the subpopulation of T4+ inducer cells responsible for maximal helper activity in T-B interactions is restricted to a minor subpopulation of T4+ lymphocytes.
Ia-dependent interleukin 2 production in syngeneic cellular interactions.
Authors: Lattime E C , Gillis S , Pecoraro G , Stutman O
Published: December 2022

In the murine syngeneic mixed lymphocyte response (SMLR), the co-culture of nylon wool-nonadherent spleen cells or young (4 to 6-day-old) thymocytes with syngeneic nylon wool-adherent adult spleen cells, results in proliferation and Interleukin 2 (IL 2) production. In both the thymus and spleen SMLR, proliferation and IL 2 production are the function of Lyt-1+2-3- (Lyt-1) responder cells and require the expression of IA (IB) and/or IE cell surface antigens on the stimulator cell. IL 2 production in both spleen and thymus SMLR is independent of heterologous serum factors (i.e., fetal calf serum), which, however, influences the magnitude of the in vitro proliferation. The IL 2-producing cell in spleen is capable of self-renewal in the absence of the thymus; it is detectable 6 to 7 mo after adult thymectomy. These results suggest the in vitro SMLR may measure a cellular interaction between the Lyt-1 subset of T cells and self Ia antigens on macrophage-like cells that produces IL 2 in an antigen-independent fashion. This interaction may have physiologic significance in the expression and regulation of T cell function.
Characterization of the alloactivated E+Ia+ cell capable of stimulating in mixed lymphocyte culture.
Authors: Shen H H , Irigoyen O , Friedman S M , Chess L
Published: December 2022

Human peripheral blood E+ lymphocytes were alloactivated by E- cells and then purified by repeat E+ selection. As described by others, alloactivated cells (E+d6) but not unactivated E+ cells were capable of stimulating both allogeneic and autologous mixed lymphocyte cultures (MLC). To further characterize the subset of cells responsible for this function, we used a variety of monoclonal antibodies to human T cells and monocytes. Treatment of E+d6 cells with anti-Ia OKI1) plus complement (C) abrogated their stimulating capacity. In contrast, treatment with the anti-T cell antibodies OKT3, OKT11A, OKT4, OKT8 plus C or anti-macrophage antibody (OKM1) plus C failed to eliminate their MLC stimulatory capacintibodies to human T cells and monocytes. Treatment of E+d6 cells with anti-Ia OKI1) plus complement (C) abrogated their stimulating capacity. In contrast, treatment with the anti-T cell antibodies OKT3, OKT11A, OKT4, OKT8 plus C or anti-macrophage antibody (OKM1) plus C failed to eliminate their MLC stimulatory capacintibodies to human T cells and monocytes. Treatment of E+d6 cells with anti-Ia OKI1) plus complement (C) abrogated their stimulating capacity. In contrast, treatment with the anti-T cell antibodies OKT3, OKT11A, OKT4, OKT8 plus C or anti-macrophage antibody (OKM1) plus C failed to eliminate their MLC stimulatory capacity. Because OKT3 recognizes the majority of T cells and OKT11A recognizes virtually all E+ cells, we reasoned that a contaminating non-T cell containing the MLC-stimulating capacity may be present within the alloactivated E+ population. To further address this question, E+d6 cells were positively and negatively selected using a rosetting technique with anti-Ig-coated red cells. The positively selected OKT3, OKT11A, OKT4, or OKT8 E+d6 cells retained minimal ability to stimulate MLC, whereas the corresponding negatively selected populations were highly enriched in this function. Phenotypic analysis of the isolated populations failed to demonstrate greater than 1% surface membrane Ig(SmIg) positive cells. Taken together, these results suggest that the MLC stimulatory capacity of alloactivated E+ cells is contained within an Ia+, OKM1-, SmIg- non-T cell subset.