bluuebunny/crossref_metadata_2025 huggingface dataset:
Stimulation of mouse lymphocytes by a mitogen derived from Mycoplasma arthritidis. III. Ir gene control of lymphocyte transformation correlates with binding of the mitogen to specific Ia-bearing cells.
Authors: Cole B C , Daynes R A , Ward J R
Published: December 2022

The supernatant from Mycoplasma arthritidis broth cultures (MAS) contains a T cell mitogen that is under Ir gene control. Responsiveness to this mitogen is dictated by the I-E/I-C subregion of the major histocompatibility complex and is dependent upon adherent radioresistant Ia+ accessory cells from responding haplotype animals. In this study, we established that MAS could be removed from culture supernatants by absorption with spleen cells from mice that themselves are responsive to the mitogen (k and d haplotypes), but activity is not removed by spleen cells from mouse strains that are nonresponsive to the mitogen (b, q, and s haplotypes). Absorption studies with lymphoid cells from congenic and recombinant strain mice established that absorption of the mitogen was itself linked to the I-E/I-C subregion of the major histocompatibility complex. Thymocytes from responding haplotype strains were incapable of removing MAS activity, and spleen cells devoid of Thy-1-positive cells retained their full absorbing capacity. The ability to effectively absorb MAS activity was abrogated by the pretreatment of spleen cells with anti-Ia antiserum and complement. Furthermore, the ability of spleen cells from responding haplotype strains to respond to MAS was blocked by the addition of anti-Ia serum to the cell cultures. Whereas the latter treatment resulted in an almost complete elimination of MAS responsiveness, the ability of similarly treated spleen cells to respond to the mitogens PHA and Con A was only minimally depressed. These results are consistent with our hypothesis that the mitogenic moiety of MAS actually binds to I-E/I-C-coded Ia antigens.
IgG induction in a human B cell line by red cell-mediated microinjection of the cytoplasm from T cell factor-stimulated B cells.
Authors: Kishimoto T , Miki Y , Kishi H , Muraguchi A , Kishimoto S , Yamamura Y
Published: December 2022

Generation of a cytoplasmic factor(s) that induced IgG secretion in nonstimulated cells was demonstrated in TRF-stimulated cells by using red cell-mediated microinjection. Injection of the cytoplasm from a TRF-stimulated B lymphoblastoid cell line (CESS) into nonstimulated cells induced an increase of IgG-producing cells. Injection of TRF itself did not induce an increase of IgG-producing cells. Active substance(s) in the cytoplasm were generated at 2 hr after TRF stimulation, and IgG-producing cells reached their maximum level at 40 hr after injection of the factor(s). DFP, but not actinomycin D, inhibited the generation of the cytoplasmic factor(s). The activity of the cytoplasmic factor(s) was not destroyed by RNase and not absorbed with anti-IgG. These results suggested that binding of TRF with its acceptors induced the generation of the cytoplasmic factor(s) involved in the transmission of TRF-mediated signals from membrane to nuclei.
Phenotypic characteristics of cell lines requiring interleukin 3 for growth.
Authors: Ihle J N , Keller J , Greenberger J S , Henderson L , Yetter R A , Morse H C
Published: December 2022

A number of cell lines have been derived from bone marrow cultures in the presence of WEHI-3 conditioned media (CM) that continue to require WEHI-3 CM for growth in vitro. Because the WEHI-3 cell line has been shown to constitutively produce a lymphokine (IL 3) that induces the expression of 20 alpha-hydroxysteroid dehydrogenase (20 alpha SDH) in cultures of splenic lymphocytes from athymic mice, we examined whether these cell lines were dependent upon IL 3 for growth. The results demonstrate that the factor required for growth of these cell lines copurifies with IL 3 activity on G-100, DEAE cellulose, and CM cellulose column chromatography as well as in preparative isoelectric focusing and on hydrophobic supports in reverse phase high pressure liquid chromatography. The biologic activity of peak fractions in each case was similar in both types of assays. These results strongly suggest that the WEHI-3 CM-dependent cell lines are dependent on IL 3 for growth in vitro. All the cell lines have readily detectable levels of 20 alpha SDH but have differing cell surface phenotypes. The C3HSFFV line is devoid of conventional lymphoid cell surface markers with the exception of Lyt-1, whereas the FDC-P1 expresses Thy-1, Ly-5, and H-11. Other cell lines have intermediate phenotypes.
Distribution of terminal deoxynucleotidyl transferase and purine degradative and synthetic enzymes in subpopulations of human thymocytes.
Authors: Ma D D , Sylwestrowicz T A , Granger S , Massaia M , Franks R , Janossy G , Hoffbrand A V
Published: December 2022

Terminal deoxynucleotidyl transferase (TdT) and purine metabolic enzymes were examined in subsets of human infant thymocytes (defined by surface cell antigens) and normal peripheral T lymphocytes. Putative prothymocytes (RFB-1+, HTA-1+/- large blast-like cells), medium and high density cortical thymocytes (RFB-1+, HTA-1+), and medullary thymocytes (RFB-1-, HTA-1-, OKT3+) were isolated by density gradient centrifugation, monoclonal antibody and complement-mediated cytolysis, and cell-antibody affinity chromatography. Peripheral T lymphocytes were isolated from normal adult mononuclear cells using nylon fiber technique. Adenosine deaminase (ADA) and TdT were highest in prothymocytes 48.8 +/- 14.7 mumol/hr/10(8) cells (mean +/- SE) and 22.9 +/- 1.4 U/10(8) cells, respectively. Both enzymes decreased progressively down the maturation pathway. In peripheral T lymphocytes, ADA was 3.9 +/- 1.5 mumol/hr/10(8) cells, and TdT was undetectable. Purine nucleoside phosphorylase (PNP) and ecto-5'nucleotidase (5'NT) were lowest in cortical thymocytes (27.5 +/- 11.0 nmol/hr/10(6) cells and 2.8 +/- 1.3 nmol/hr/10(6) cells, respectively) and increased with T cell maturation. The PNP level was 124.9 +/- 17.2 nmol/hr/10(6) cells and 5'NT was 30.1 +/- 3.9 nmol/hr/10(6) cells in peripheral T lymphocytes. The deoxynucleoside kinases (deoxyguanosine, deoxyadenosine, and deoxycytidine kinases) paralleled the changes in ADA and TdT activity among the different T subsets. The proliferative activity (labeling index) was highest in the prothymocyte fraction and lowest in peripheral T cells. Variation in the distribution of these enzymes in T cell subsets may explain their different sensitivities to deoxyadenosine and deoxyguanosine toxicity and the different effects on T cell development of ADA or PNP deficiency.
Corticosteroid alteration of human monocyte to macrophage differentiation.
Authors: Rinehart J J , Wuest D , Ackerman G A
Published: December 2022

Human monocyte to macrophage differentiation in vitro is associated morphologically with an increased cell size, an increased number of lysosomes, and rough endoplasmic reticulum. Functional changes associated with monocyte to macrophage differentiation include increased tumoricidal activity and increased cell protein, acid phosphatase, and 5'-nucleotidase. Hydrocortisone succinate (HCS) at 2.5 microM markedly altered monocyte to macrophage differentiation: HCS inhibited the development of tumoricidal activity and the increased levels of cell protein, acid phosphatase, and 5'-nucleotidase. By transmission electron microscopy, macrophages incubated with HCS failed to develop an increased complement of lysosomes and developed an increased number of membrane-bound electron lucent vacuoles. Dexamethasone inhibited the development of tumoricidal activity at a 10-fold lower concentration than HCS. HCS also markedly inhibited monocyte 3H-uridine incorporation. Mechanisms of HCS alteration of monocyte differentiation are discussed. These data suggest that corticosteroid alteration of monocyte differentiation may be a mechanism of HCS immunosuppression in vivo.
Two monoclonal anti-human T lymphocyte antibodies have similar biologic effects and recognize the same cell surface antigen.
Authors: Burns G F , Boyd A W , Beverley P C
Published: December 2022

Two independently produced monoclonal antibodies, UCHT1, an IgG1 antibody, and OKT3, an IgG2 antibody, which specifically identify all human peripheral T lymphocytes, were shown to possess similar functional properties. Both were highly mitogenic to peripheral blood T lymphocytes, and UCHT1 blocked specific T lymphocyte killing of autologous B lymphoblasts when present during either the induction or the cytotoxic phase of the reaction; this, too, is a property reported for OKT3 in an analogous situation. Both antibodies also inhibited the 6-day proliferative response to soluble antigens and mixed lymphocyte culture, although this may be attributable to their strong mitogenicity earlier in culture. These properties were not found with other monoclonal antibodies directed against human T lymphocytes, and suggested that UCHT1 and OKT3 recognize important functional structure(s) on the T cell surface. The identity of the antigen recognized by these two antibodies was demonstrated by 2-directional co-capping, by cold antibody inhibition of binding of radiolabeled antibody, and by immunoprecipitation and two-dimensional gel analysis of labeled T cell surface proteins. Both UCHT1 and OKT3 bound with similar affinities (Ka approximately 2 X 10(9) M-1) to the same number of binding sites (2.8 X 10(4) sites/activated T cell), and both precipitated the same glycoprotein complex with a m.w. of approximately 19,000 in the presence of reducing agent.
Monoclonal antibodies to streptococcal group A carbohydrate. I. A dominant idiotypic determinant is located on Vk.
Authors: Nahm M H , Clevinger B L , Davie J M
Published: December 2022

In an effort to better define the antibody repertoire to streptococcal group A carbohydrate (GAC), somatic cell hybrids were prepared from A/J mice immunized with streptococcal vaccine. Most antibodies were IgG3K and IgMK, while 2 of 26 antibodies were lambda type. Each of the IgG3 antibodies had a distinct isoelectric point consistent with previous estimates of clonal repertoires of approximately 200. IEF analysis of the L chains, however, showed that about half of the antibodies produce a common L chain, called VK1GAC, previously identified in A/J anti-GAC serum antibodies. Additional support for the structural similarity of these L chains was gained by developing an idiotype antiserum to VK1GAC. All proteins with the common L chain spectrotype react strongly with anti-VK1GAC. Thus, it appears that anti-GAC antibodies are composed of H chains bearing a few VH regions pairing with a few L chains.
Prostaglandin D2 generation after activation of rat and human mast cells with anti-IgE.
Authors: Lewis R A , Soter N A , Diamond P T , Austen K F , Oates J A , Roberts L J
Published: December 2022

Anti-IgE-dependent activation of rat and human mast cells resulted in the preferential generation of the cyclooxygenase products prostaglandin D2 (PGD2) and prostaglandin I2 (PGI2) in the rat and PGD2 in the human. The average net generation of PGD2, determined by gas chromatography-mass spectrometry, was 13.1 ng/10(6) purified rat mast cells and 39.5 ng/10(6) dispersed, enriched human mast cells. After IgE-dependent activation, there was a linear relationship between the net quantities of PGD2 generated and of histamine secreted from dispersed human pulmonary cells when the number of mast cells was varied but the total number of cells was held constant, indicating that it is the number of mast cells participating in IgE-dependent activation, rather than total mast cell number, that determines PGD2 generation. A linear relationship was also shown between PGD2 generation, determined by radioimmunoassay, and the release of the granule marker beta-hexosaminidase from purified rat mast cells on the dose-response portion of the plot of their response to anti-IgE challenge. With higher concentrations of anti-IgE, PGD2 generation from rat mast cells plateaued, whereas net percent beta-hexosaminidase release increased further. In kinetic studies of rat mast cells activated with anti-IgE, the onset (1 to 2 min) and time of maximum generation (5 to 10 min) for PGD2 were delayed relative to the onset (15 to 30 sec) and completion (1 to 2 min) of beta-hexosaminidase release. Thus, the extracellular appearance of PGD2 during IgE-dependent mast cell activation represents a response additional to the secretion of granule-associated mediators.
Identification of T cell subsets and B lymphocytes in mouse brain experimental allergic encephalitis lesions.
Authors: Sriram S , Solomon D , Rouse R V , Steinman L
Published: December 2022

Tissue section immunoperoxidase staining, employing monoclonal antibodies specific for T cell antigens (Lyt-1, Lyt-2) and B cells (B220), was used to study the brains of mice with experimental allergic encephalitis. Cells bearing phenotypes characteristic of both helper (Lyt-1+2-) and cytotoxic/suppressor (or precursor) (Lyt-2+) T cells, as well as B cells, are present in the perivascular cuffs in brains of EAE mice during active disease and the recovery phase. Lyt-1+ cells comprise 49% of the inflammatory cell population, and Lyt-2+ and B220+ cells are found at frequencies of 10% and 12%, respectively.
The immune response to homologous lens crystallin. I. Antibody production after lens injury.
Authors: Goldschmidt L , Goldbaum M , Walker S M , Weigle W O
Published: December 2022

Using a sensitive radioimmunoassay to homologous mouse alpha-crystallin, it was established that autoantibodies are produced to this self constituent in mice after rupture of the lens capsule by needling. Antibody to lens crystallin was detected within 4 days of lens rupture and persisted for 24 days, suggesting that these mice are not tolerant to alpha-crystallin at the B cell level. However, T cells from mice immunized with homologous crystallin cannot be stimulated to proliferate by either homologous or heterologous crystallin. On the other hand, lymphocytes from mice immunized with bovine alpha-crystallin can be stimulated to proliferate to bovine crystallin, but not to either unfractionated mouse crystallin (MC) or purified mouse alpha-crystallin. At the level of detection, therefore, T cells appear to be tolerant to homologous alpha-crystallin, but not to heterologous alpha-crystallin. LPS resulting from bacterial contamination after needling was ruled out as a necessity for antibody production because antibody to mouse alpha-crystallin was produced after sterile lens rupture with a laser. However, LPS and poly A:U did enhance the antibody response to alpha-crystallin. These data suggest that T cells, but not B cells, are tolerant to homologous crystallin and that antibody results from circumvention of specific helper T cell requirements for antibody synthesis. The implications of these findings for tolerance and autoimmune uveitis are discussed.
Enhanced resistance to Nocardia brasiliensis infection in mice depleted of antigen-specific B cells.
Authors: Rico G , Ochoa R , Oliva A , Gonzalez-Mendoza A , Walker S M , Ortiz-Ortiz L
Published: December 2022

This study deals with the roles of T and B cells in Nocardia brasiliensis infection in mice. Nocardia injected into the animals' footpads caused inflammatory responses and mycetomas in situ, resulting in granulomatous lesions of subcutaneous tissues and eventual bone destruction. These clinical features resemble those of humans infected with Nocardia. The effect, if any, of antibody was studied by passively transferring anti-Nocardia serum into either immunologically normal or T-deficient infected mice. Such transfers had no protective function in either group. To the contrary, the antibody seemed to favor infection and worsen bone disease compared to that in mice not given antibody. Furthermore, passive transfer of the antibody along with injection of Nocardia coated with the antibody magnified the severity of subsequent symptoms. Although these experiments ruled out any role for antibody in protection from Nocardia, they did not directly prove T cell participation in such resistance. Therefore, the role of T cells during Nocardia infection was examined further by transferring spleen cells depleted of B lymphocytes bearing receptors for a Nocardia extract (NE). Lethally irradiated mice reconstituted with a population depleted of NE-specific B cells totally lacked the ability to form antibodies to NE; however, they mounted effective delayed-type hypersensitivity reactions and completely controlled their Nocardia infection, establishing the importance of cell-mediated immunity in halting this disease process.
Cloned lines of human cytotoxic T lymphocytes with specificity for influenza virus.
Authors: Fleischer B
Published: December 2022

The generation of human cytotoxic T cell clones with specificity for influenza virus and some of their characteristics are described. The clones were generated by limiting dilution of peripheral blood lymphocytes after two in vitro stimulations with autologous influenza A/USSR virus-infected cells and were grown in T cell growth factor. The majority of the virus-specific clones showed cross-reactivity for different influenza A virus subtypes but did not recognize influenza B virus-infected cells. The HLA specificity of two clones was further analyzed. One clone, LL33, was specific for HLA-Bw60, the other, clone WH5, for HLA-A1. Clone WH5 also seemed to recognize the serologically related HLA-A26 as restriction element for the recognition of the viral antigen. Whereas the virus-specific CTL clones had the OKT3+,4-,8+ phenotype, another clone, WH 49, exhibiting natural killer-like activity, was found to have the OKT3+,4+,8- phenotype.
Depressed T cell proliferative responses in Hodgkin's disease: role of monocyte-mediated suppression via prostaglandins and hydrogen peroxide.
Authors: Fisher R I , Bostick-Bruton F
Published: December 2022

We studied the contribution and mechanisms of monocyte-mediated suppression in the depressed T cell proliferative responses observed in patients with Hodgkin's disease. Mononuclear leukocytes from 22 untreated patients had significantly lower proliferation after stimulation with suboptimal doses of phytohemagglutinin than normals controls (p less than 0.001). When indomethacin was added to the cultures, the mean percent increase in patient proliferation exceeded that of normals (160 +/- 19% vs 81 +/- 9%; p less than 0.008), yet patient proliferation remained only 36% of normal. Catalase alone caused a minimal increase in proliferation in all cultures. When catalase plus indomethacin were added to either normal or patient cultures, however, a synergistic increase was observed. The mean percent increase in patient proliferation was 300 +/- 80%, although once again the absolute proliferative response of the patients remained subnormal (p less than 0.003). Removal of adherent monocytes from patient cultures produced increases in proliferation comparable to that observed with the addition of indomethacin. Long-term disease-free survivors of Hodgkin's disease had depressed T cell proliferation but no significant increase after the addition of indomethacin. We conclude: 1) although untreated patients with Hodgkin's disease have increased monocyte suppressor activity that is mediated by increased prostaglandin production, this is not the major cause of the depressed T cell proliferative responses observed in Hodgkin's disease; 2) depressed proliferative responses in cured Hodgkin's disease are not mediated by prostaglandins; 3) hydrogen peroxide suppresses T cell proliferation in both normals and untreated patients; and 4) other factors, probably inherent in the T cell itself, are the major cause of depressed T cell responses in Hodgkin's disease.
Characterization of the cytolytic reaction mechanism of the human natural killer (NK) lymphocyte: resolution into binding, programming, and killer cell-independent steps.
Authors: Hiserodt J C , Britvan L J , Targan S R
Published: December 2022

Various parameters of the cytolytic reaction mechanisms of the human natural killer (NK) lymphocyte were studied to characterize the lytic cycle. NK cytolysis was determined to occur in three definable steps. 1) Binding of PBL to the NK-sensitive targets Molt-4 or K562 was rapid (less than 1 min), occurred at temperatures below 37 degrees C, was Mg++3-dependent, Ca++3-independent, and was prevented by dispersion of the cells into 10% dextran. 2) Subsequent to binding, programming for lysis as determined by a Ca++ pulse method was more protracted, requiring up to 2 hr to occur and was strictly dependent on Ca++ for cytolysis to proceed. In standard cytotoxicity assays, however, programming for lysis was more rapid occurring in 10 to 30 min. Programming was inhibited by EDTA, EGTA/Mg++ and by temperatures below 37 degrees C. Furthermore, after binding but in the absence of initiation of programming for lysis, the frequency of target binding cells did not change and the NK cell did not lose its lytic potential. 3) Killer cell-independent cytolysis (KCIL) was determined by the addition of EDTA to "programmed" targets and dispersion of these cells into dextran-containing medium, which resulted in virtually 100% dissociation of conjugated cells. KCIL was Ca++ and Mg++-independent and was blocked at reduced temperatures only if the dextran was prechilled to 4 degrees C before addition. The kinetics of 51Cr release during KCIL was rapid and complete 30 min after dispersion. Interferon-activated NK cells expressed an increased rate of cytolysis in Ca++ pulse experiments. This was due to an increased rate of the Ca++-dependent step(s) during the programming events. The rate of the Ca++-independent steps, however, were similar with control and IFN-activated cells.
Abnormalities induced by the mutant gene Ipr: expansion of a unique lymphocyte subset.
Authors: Morse H C , Davidson W F , Yetter R A , Murphy E D , Roths J B , Coffman R L
Published: December 2022

Mice carrying the Ipr mutation develop massive lymphoadenopathy and severe autoimmune disease. The characteristics of the cell population that proliferates in lymphoid tissues were evaluated by the use of a) monoclonal antibodies and FMF, and b) molecular genetic studies of Ig heavy chain genes. The lymph node cells of different strains of mice homozygous for the Ipr mutation were shown to be almost uniformly Thy-1+, Ly-1+, Ly-2-, H-11+, Ly-5+, sIg-, ThB-, 2C2+, I-A-, 6B2+, and therefore to have surface characteristics of both T and B cells. Molecular genetic studies of the arrangements of Ig heavy chain genes showed that they were not rearranged as in pre-B and B cells. These results suggest that an abnormal proliferating population of T cells in Ipr/Ipr mice aberrantly express B cell surface markers.
Dexamethasone induces different cellular protein synthetic responses in PNA+ and PNA- mouse thymocyte subpopulations.
Authors: Khalid B A , Pearce P , Barr I G , Fraillon D , Toh B H , Funder J W
Published: December 2022

Thymocytes from adrenalectomized BALB/c male mice were separated by peanut agglutination (PNA) into cortical, corticosensitive, PNA+ cells and larger, medullary, corticoresistant, PNA- cells; the extent of cross-contamination of PNA+ and PNA- cells, and vice versa, was checked by flow microfluorometry. Glucocorticoid receptor profiles were established with 3H-dexamethasone as probe; no differences in receptor affinity or cellular concentration, or in cytoplasmic and nuclear compartmentalization were seen between PNA+ and PNA- cells. On two-dimensional gel electrophoresis, PNA+ and PNA- thymocytes from oil-injected (control) adrenalectomized mice showed patterns of incorporation of 35S-methionine into protein that differed in at least 12 spots, as revealed by autoradiography. PNA+ and PNA- cells from mice treated with submaximal (6 micrograms/day) or near-maximal thymolytic doses of dexamethasone (20 micrograms/day) were also examined by two-dimensional gel electrophoresis. Both PNA+ and PNA- cells showed substantial, overlapping dexamethasone-induced changes in protein synthetic profiles.
C-reactive protein- (CRP) mediated modulation of human B cell colony development.
Authors: Whisler R L , Newhouse Y G , Mortensen R F
Published: December 2022

The ability of human C-reactive protein (CRP) to modulate Staph protein A-(SpA) induced human B cell colony formation in semisolid cultures was investigated. Maximal augmentation was observed with 10 and 25 micrograms/ml CRP and was independent of the SpA concentration used to stimulate colony formation. Optimal facilitation of colony numbers was noted when CRP was present during the initial stages of colony formation and the facilitation represented an early increase in colony numbers. At later incubation intervals, however, the colony responses of cultures containing 25 and 50 micrograms/ml CRP were considerably diminished when compared to controls that appeared secondary to the focal disintegration of colony clusters rather than the actual suppression of certain colony progenitors with delayed colony-forming kinetics. Other experiments showed the 18-hr pre-exposure of cells to CRP facilitated colony formation, and that after preincubation, a small frequency of CRP binding cells could be detected by flow microcytofluorometry. Therefore, these studies indicate that a possible biologic function of CRP is the ability to modulate the activities of the B cell system.
Release of a lymphokine-like plasminogen activator by stimulated B lymphocytes.
Authors: Toullet F , Maillard J L , Favreau C
Published: December 2022

This report extends to the guinea pig the discovery of a lymphocyte plasminogen activator (LPA) previously described in the mouse. In the guinea pig, we have identified enzymatic activity similar to that in the mouse, but which has two distinct components: first, as in the mouse, a membrane-bound molecule present even in the quiescent lymphocyte (mLPA), and second, a previously unreported soluble molecule appearing in the culture medium after appropriate cell stimulation (sLPA). This sLPA, like most known lymphokines, was released by in vitro recall of sensitized lymphocytes by the antigen and by direct contact with a mitogen. There was a parallel evolution in the culture for sLPA and for some other well-known lymphokines (LT, LIF), and their detection thresholds were of the same order. A possible activation of plasminogen (Pg) by macrophages contaminating the lymphocyte cultures was carefully rule out. sLPA was produced by lymph node lymphocytes as well as by blood lymphocytes, but not by spleen cells, thymocytes, or peritoneal lymphocytes. A study of the kinetics of the release of sLPA, together with that of metabolic modifiers, suggested that an intracellular synthesis precedes the secretion of the molecule. Data obtained from the use of B- and T-enriched subpopulations or B- or T-dependent antigens and mitogens point to the B lymphocytes as the major, if not exclusive, source of sLPA. The choice of synthetic chromogenic substrates S 2251, S 2444, and S 2288 in some experiments led us to confirm most of the above properties of sLPA with still greater precision and reliability.
Analysis of the oligosaccharides on the HLA-DR and DC1 B cell antigens.
Authors: Shackelford D A , Strominger J L
Published: December 2022

The types of N-linked oligosaccharides on the HLA-DR and DC1 antigens were determined by their sensitivity to endoglycosidase H or endoglycosidase D. The DR and DC1 heavy chains each have two carbohydrate moieties, one high-mannose and one complex-type glycan. The DR and DC1 light chains have one complex-type oligosaccharide. During the biosynthesis of the DR antigens, the oligosaccharides on both subunits are initially high-mannose-type glycans, as has been found for other membrane glycoproteins. The light chain oligosaccharide and one of the two heavy chain carbohydrates are later processed to complex-type glycans. Inhibition of N-linked glycosylation with tunicamycin does not inhibit chain association of the DR and DC1 subunits, transport to the cell surface, or expression of the alloantigenic determinants.
Stable hamster-mouse hybridomas producing IgG and IgM hamster monoclonal antibodies of defined specificity.
Authors: Sanchez-Madrid F , Szklut P , Springer T A
Published: December 2022

Specific antibody-secreting hybridomas have been obtained by fusing Syrian or Armenian hamster (Mesocricetus auratus or Cricetulus migratorius) spleen cells with mouse myeloma cells. The hamsters were immunized to mouse cytolytic T lymphocytes. Hybrids were selected either by an indirect binding assay using an 125I-monoclonal antibody (MAb) reactive with hamster kappa-chains or by their ability to block T cell-mediated cytolysis. Three hybridoma clones were obtained that secreted intact IgM-like and IgG-like hamster MAb as shown by SDS-PAGE. The clones were stable as shown by subcloning. Two MAb recognized antigens of wide tissue distribution; the third bound specifically to T lymphocytes, gave strong inhibition of T cell-mediated cytolysis, and immunoprecipitated the Lyt-2,3 molecule.
Biochemical evidence for expression of a semi-allogeneic, H-2 antigen by a murine adenocarcinoma.
Authors: Callahan G N , Pardi D , Giedlin M A , Allison J P , Morizot D M , Martin W J
Published: December 2022

LT-85 is an alveolegenic adenocarcinoma induced in mutant C3HfB/HeN (C3Hf) mice. This tumor, however, grows preferentially in allogeneic, wild-type C3H/HeN (C3H) mice. The tumor-associated transplantation antigen has been mapped to the K end of the major histocompatibility complex. H-2K antigens were isolated from detergent extracts of LT-85 cells by immunoprecipitation with monoclonal antibody. The tryptic peptides of these antigens were compared, by using high-pressure liquid chromatography, with the tryptic peptides of H-2K antigens isolated from syngeneic mutant C3Hf and ancestral wild-type C3H spleen cells. We found that the H-2K antigens of the LT-85 tumor cells were very similar to, but distinct from, those present on syngeneic C3Hf lymphoid cells. We also found, however, that the H-2K antigens of LT-85 tumor cells were clearly different from the H-2K antigens of allogeneic C3H spleen cells. The H-2K antigens of LT-85 cells are therefore foreign to syngeneic C3Hf cells, but do not represent expression by the tumor cells of the allogeneic H-2K antigens expressed by normal C3H cells. Furthermore, the nature of the differences observed between the H-2K antigens of LT-85 cells and C3Hf and C3H spleen cells strongly suggests that the structure of the H-2K molecule of LT-85 cells is identical in some regions to the H-2K molecule of C3Hf cells, and in other regions to the H-2K molecule of C3H cells.
Evidence for cytostatic T cell activity in the effector mechanism against syngeneic TMT mammary tumor cells in mice.
Authors: Kurata S , Tsuchiya T , Norimura T , Yamashita U
Published: December 2022

The effector mechanism of immune spleen cells against syngeneic TMT mammary tumor cells was analyzed in vitro. C3H/He mice were first inoculated with TMT tumor cells, and then the tumors were x-irradiated with 2000 rad 1 wk after the inoculation. Spleen cells from these treated mice inhibited the growth of tumor cells in vitro when assessed by (3H)-TdR incorporation by tumor cells (cytostatic activity). The same spleen cells did not have any cytotoxic activity on TMT tumor cells detected by a 51Cr-release assay. The cytostatic activity was mediated by Lyt-1+23- T cells. The purified T cells alone could not inhibit the growth of tumor cells, but accessory cells were required for the induction of cytostatic T cell activity. The accessory cells were Ia-positive, macrophage-like adherent cells. Furthermore, both T cells and macrophages were also required for the inhibition of tumor growth even after the spleen cells were activated in vitro. These results suggest T cells and macrophages play an important role in the effector mechanism against TMT mammary tumor cells. The mechanism of cytostasis by T cells and macrophages was discussed from the standpoint of the cellular interaction.
Immunologic suppression of carcinogenesis in spontaneously hypertensive rats (SHR) with T cell depression.
Authors: Takeichi N , Ba D , Koga Y , Kobayashi H
Published: December 2022

A strain of spontaneously hypertensive rats (SHR) showed a selective depression of T cell functions brought about by aging. Conversely, this strain had a high NK cell activity as compared to other normal rat strains. This SHR strain was found to be much more sensitive to the carcinogenic activity of low doses of MCA than were WKA rats with normal T cell functions. Allogeneic thymus grafts almost completely restored the T cell functions of SHR, whereas injection of an immunopotentiator, NSP, enhanced NK cell activity and also caused a partial recovering of T cell functions. When immunologic restoration was achieved, generation of killer T cells to syngeneic SMT-5 tumor cells was induced and the cytotoxic activity of NK cells to K-562 cells was also enhanced. But the cytotoxic activity to the SMT-5 cells of NK cells and macrophages from the treated or untreated SHR was not detected. Allogeneic thymus grafts induced a significant transplantation resistance against a syngeneic SMT-5 tumor and injection of NSP enhanced only the survival days of the rats. Allogeneic thymus grafts also significantly suppressed the incidence of tumors induced by MCA, whereas the injection of NSP was not effective in the prevention of tumor development but was effective in prolongation of latency periods. These results support the hypothesis that immune surveillance mediated by T cells is an important mechanism for the control of tumor development.
Fine-specificity of the contact sensitivity to 4-hydroxy-3-nitrophenyl acetyl (NP).
Authors: Matoso-Ferreira A , Mäkelä O
Published: December 2022

Dorf and colleagues (1-4) found that the contact sensitivity (CS) primed with (4-hydroxy-3-nitrophenyl)acetyl (NP) could be elicited as easily with the iodoanalog (NIP) as with NP when studied in Igh-1b mice but could only be elicited with NP, not NIP, in Igh-1j mice. Since this fine-specificity was parallel to the fine-specificity of anti-NP antibodies in the two types of mice and since anti-NP antibodies of Igh-1b mice are controlled by gene Igh-NPb the authors concluded that CS also was controlled by the Igh-NPb gene. The aim of this study was to confirm their findings with a more quantitative method (5). We confirmed equality of NP and NIP as elicitors of NP-primed CS in Igh-1b mice when the priming antigen was given subcutaneously into non-cyclophosphamide-treated mice (their method). We also found that this priming induced an anti-NP antibody response detectable at the time of challenge. Most experiments were carried out with a method that does not induce a detectable antibody response (pretreatment of mice with 200 mg/kg of cyclophosphamide; application of the sensitizing compound on skin). Since the NP-primed (and NBrP-primed) CS reactions exhibited "expected specificities," the immunizing compound was clearly the most efficient elicitor (relative efficiencies of homologs varied from 2 to 4). The Igh-NPb gene appears not to have a role in "antibody-free" reactions.
Immunoglobulin idiotype and anti-anti-idiotype utilize the same variable region genes irrespective of antigen specificity.
Authors: Margolies M N , Wysocki L J , Sato V L
Published: December 2022

Idiotypic determinants characterizing certain antibody specificities have been proven valuable structural and genetic markers in studies of antibody diversity and regulation. The heritable predominant idiotype associated with the response to p-azophenylarsonate in A/J mice consists of a set of highly homologous (greater than 95%) heavy and light chain variable region amino acid sequences probably arising by somatic mutation from one or a few V region genes. We examined a peculiar set of monoclonal antibodies that have been defined as CRI by serologic analysis, but that have no affinity for the hapten Ars. These antibodies were elicited by immunization with anti-CRI rather than by the conventional immunization with antigen. The amino acid sequences of the amino terminal half of the V regions of these anti-(anti-CRI) antibodies are indistinguishable from those of conventional Ars-binding CRI antibodies. Thus, Ars-binding CRI and Ars-nonbinding anti-(anti-CRI) are derived from similar or identical VH and VL genes.
Helper cell-independent cytolytic T lymphocytes specific for a minor histocompatibility antigen.
Authors: Roopenian D C , Widmer M B , Orosz C G , Bach F H
Published: December 2022

We have isolated several anti-H-2Db-restricted cytolytic T cell clones specific for the minor histocompatibility antigen, H-1, from a long-term B10.129(5M) anti-C57BL/6 mixed leukocyte culture. All proliferate significantly, in the absence of exogenously added lymphokine, and in a specific manner to H-2Db-compatible stimulator cells that carry the appropriate H-1 minor antigen. Moreover, significant proliferation also occurs when T cell-depleted stimulator cells are used as the antigenic stimulus, indicating that the proliferative capacity of these clones is not the result of T cell factors produced by the stimulator cells. After concanavalin A stimulation, IL 2-like lymphokine activity can be detected from the one clone tested, 5M/B6-5. These data demonstrate the existence of antigen-driven, helper T cell-independent cytolytic T cell clones responsive to a single minor histocompatibility antigen. The implications of this finding are discussed.
Mechanisms of idiotype suppression. V. Anti-idiotype antibody inhibits early B cell triggering induced by antigen.
Authors: Kim B S , Massey P B , Hui K M
Published: December 2022

To investigate the relationship between antigen-mediated B cell commitment and induction of idiotype (id) suppression, anti-id antibody directed against the major id (TEPC-15 idiotype or T15id) of the anti-phosphorylcholine (PC) antibody was added at various time intervals to BALB/c spleen cell cultures stimulated with a T-independent PC antigen, R36a. The suppressive effect of anti-T15id antibody on the anti-PC response was rapidly decreased as addition of the antibody was delayed; when anti-T15id antibody was added 6 hr after the initiation of the cultures, only partial suppression was induced, whereas the addition of anti-id antibody after 24 hr did not result in significant suppression of the anti-PC response when compared with similar cultures treated with mock anti-id antibody. This acquisition of resistance to id suppression was completely inhibited by treatment with either sodium azide or colchicine, as well as at temperatures below 20 degrees C. The induction of resistance to id suppression during the preincubation period was dependent on the presence of an immunogenic level of specific antigen. This antigen-mediated B cell commitment did not appear to require macrophages because preincubation of macrophages with antigen did not affect the sensitivity of the B cells to anti-id antibody. These results support the possibility that anti-id antibody inhibits early B cell triggering, which involves an energy-dependent, epitope-mediated, lateral mobility of antigen receptors possibly followed by repolymerization of microtubules.
The ontogeny and distribution of B cells in normal and mutant immune-defective CBA/N mice: two-parameter analysis of surface IgM and IgD.
Authors: Scher I , Titus J A , Finkelman F D
Published: December 2022

A dual-laser fluorescence-activated cell sorter was utilized to study the distribution of the surface IgM and IgD on individual B cells of normal and immune-defective CBA/N mice. Cells from different lymphoid organs and from developing mice were studied. Two major populations of cells were seen. Those with low densities of surface IgM and intermediate-high densities of surface IgD were relatively or totally absent from the bone marrow, spleens, and lymph nodes of adult, immune-defective (CBA/N x DBA/2)F1 male mice, and developed late in ontogeny in the lymphoid organs of normal F1 female mice. By contrast, the second major population, with intermediate-high surface IgM and low surface IgD, was found in highest frequency in the lymphoid organs of immature mice, the bone marrow of adult mice, and the lymphoid organs of F1 male mice compared to F1 female mice at any age. These two major populations of B cells were further subdivided into five groups of cells to better define the surface IgM and IgD characteristics of developing B cells of immune-defective and normal mice. The relationship of these groups of cells to populations defined by other criteria are discussed.
Phenotypic characteristics of peripheral blood T cells regulating colony growth by nontransformed human B lymphocytes.
Authors: Whisler R L , Wajda K J , Newhouse Y G
Published: December 2022

The phenotypic characteristics of peripheral blood T cell subpopulations regulating human B cell colony growth stimulated by Staph protein A were investigated. Colony growth was facilitated by OKT4 cells, and T cells expressing DR antigens were found to be partially responsible for colony facilitation. A linear increase in the magnitude of colony growth was observed with greater T cell numbers, and maximal colony enhancement occurred when T cells were present during the early stages of colony formation. OKT8 cells did not enhance colony growth and also inhibited the facilitation of colony formation by OKT4 cells. Other experiments showed that the functional activities of OKT4 and OKT8 cells differed in their requirements for DNA synthesis. Although active T cell DNA synthesis was absolutely required for the facilitation of colony growth at all concentrations tested, DNA synthesis was not needed for OKT8 inhibition of OKT4 promotion of colony formation. Thus, distinct T cell subsets whose functional properties differ in their requirements for DNA synthesis regulate human colony growth.
IgG subclass expression by human B lymphocytes and plasma cells: B lymphocytes precommitted to IgG subclass can be preferentially induced by polyclonal mitogens with T cell help.
Authors: Mayumi M , Kuritani T , Kubagawa H , Cooper M D
Published: December 2022

The human IgG subclasses expressed by circulating B lymphocytes, tissue plasma cells, and plasma cells generated from B cell precursors in response to the polyclonal mitogens LPS and PWM were examined by immunofluorescence using subclass-specific monoclonal antibodies. The subclass distribution observed for circulating B lymphocytes was IgG2 (48%) greater than IgG1 (40%) greater than IgG3 (8%) greater than IgG4 (1%), while the distribution among IgG plasma cells in bone marrow, blood, spleen, and tonsils was IgG1 (64%) greater than IgG2 (26%) greater than IgG3 (8%) greater than IgG4 (1%). Multiple IgG isotypes were not observed on B cells or in plasma cells. Although IgG plasma cell responses to both LPS and PWM were T cell dependent, the distributions of IgG subclasses elicited were strikingly different. In control and LPS-stimulated cultures of blood mononuclear cells, the induced plasma cells expressed the IgG subclass distribution: IgG2 greater than 80%, IgG1 less than 20%, IgG3 less than 1%, IgG4 less than 1%. In PWM-stimulated cultures, the subclass distribution, IgG1 approximately 65%, IgG2 approximately 25%, IgG3 approximately 7%, IgG4 approximately 1%, was in perfect concordance with the in vivo subclass distribution of IgG plasma cells. Selective inhibition of suppressor T cell activity by x-irradiation and mitomycin C treatment did not alter the IgG subclass distribution pattern induced by LPS and PWM. Monoclonal antibodies were used to deplete selectively the B cell precursors bearing IgG1, IgG2, or IgG3 before PWM stimulation of blood mononuclear cells. In each instance, a reduction was observed only in the subpopulation of plasma cells producing the homologous IgG subclass. The results indicate that T cells can preferentially influence the terminal differentiation of B cells that are precommitted to different IgG subclasses.
Granulocyte activation by endotoxin. I. Correlation between adherence and other granulocyte functions, and role of endotoxin structure on biologic activity.
Authors: Dahinden C , Galanos C , Fehr J
Published: December 2022

Reminiscent of high concentrations of formylated chemotactic peptides, another group of bacteria-derived products, the lipopolysaccharides and lipid A, stimulate PMN adherence to petri dishes. Attachment and spreading of PMN is accompanied by intense release of secondary granule constituents and marked stimulation of the hexose monophosphate shunt activity. Dose-response studies with endotoxin preparations of diverse activity show that induction of PMN adherence, enzyme release, and respiratory burst activation are highly correlated, suggesting that this functional triad is mediated by a common mechanism. Hyperadhesiveness inducing concentrations of the chemotactically inert endotoxin lead to marked inhibition of PMN migration without affecting the direction-finding mechanism of the cell toward formylated peptides and C-derived chemotaxin(s). Endotoxin preparations at a lower grade of aggregation are more active, and the polysaccharide chains of the molecule are not essential with respect to PMN stimulation. Under our experimental conditions, endotoxin-induced stimulation of PMN is not inhibited by indomethacin, suggesting independence of cyclooxygenase-derived products. This type of PMN activation may play an important role in endotoxin-mediated tissue damage in vivo. Furthermore, hyperadhesion-induced inhibition of PMN migration to inflammatory sites during endotoxemia might hamper host resistance.
Acute-phase reactants of mice. II. Strain dependence of serum amyloid P-component (SAP) levels and response to inflammation.
Authors: Mortensen R F , Beisel K , Zeleznik N J , Le P T
Published: December 2022

Serum amyloid P-component (SAP) is an acute-phase reactant of mice that occurs at baseline concentrations that vary over a 10-fold range among several common inbred mouse strains. The endogenous SAP levels of all strains increased to 180 to 230 micrograms/ml 24 h after an inflammatory stimulus of either thioglycollate (TG) or lipopolysaccharide (LPS). The C57BL/6J and C57BL/10SN strains displayed 10-fold increases in SAP concentration during acute inflammation, whereas the A/J and DBA/2J strains increased levels by only 20 to 40%. These strains were therefore designated high and low SAP responders, respectively. Testing for H-2 linkage of endogenous SAP levels by using several H-2 congenic lines derived from the C57BL/10, C3H, A, and BALB parental backgrounds indicated that the levels were only weakly influenced by the H-2 complex and were influenced much more strongly by the non-H-2 background genes. F1 hybrids (B6AF1 and B6D2F1) of high and low responder strains displayed a high SAP response similar to that of C57BL/6J mice. Because IL 1-containing macrophage supernatants and semi-purified human IL 1 were previously shown by us to induce SAP in vivo, we titrated IL 1 as LAF activity in supernatants from LPS-activated macrophages from the different mouse strains. Macrophages from the high SAP responder strains displayed significantly higher IL 1 activity than macrophages from the low SAP responder strains. LPS-unresponsive C3H/HeJ or C57BL/10Sc mice did not respond to LPS with an increase in SAP synthesis. Adoptive transfer of the SAP response to C57BL 10Sc mice by using early (90 min) acute-phase sera from the different strains indicated that higher SAP inducer activity was present in the latent phase sera from the high SAP responder strains. The identification of high and low SAP responder strains should aid in determining the biologic role of this acute-phase reactant.