bluuebunny/crossref_metadata_2025 huggingface dataset:
Specific cytolysis of fresh tumor cells by an autologous killer T cell line derived from an adult T cell leukemia/lymphoma patient.
Authors: Kannagi M , Sugamura K , Kinoshita K , Uchino H , Hinuma Y
Published: December 2022

Cytotoxic T cells (Tc) derived from one patient with adult T cell leukemia/lymphoma (ATL) killed fresh autologous lymphoma cells in vitro. The Tc were induced from peripheral blood leukocytes (PBL) of this patient during remission by multiple in vitro stimulations with an autologous ATLV-bearing cell line (ILT) that was previously established by cloning of PBL in the presence of interleukin 2 (IL 2). PBL from eight other ATL patients were stimulated in the same manner, and responder cells from a patient in remission also showed cytotoxicity specific for ATL virus (ATLV)-bearing cells. Fresh lymphoma cells were obtained in relapse and were used as target cells for the autologous Tc induced. They became susceptible to the Tc within 4 hr of in vitro incubation, and their susceptibility increased with incubation time for at least 12 hr. ATLV antigens on the cell surface of these lymphoma cells, however, were not detected by radioimmunoassay during these incubation periods, but were detectable after 16 hr of incubation. In addition, cytotoxicity against lymphoma cells was completely inhibited by autologous ILT cells used as "cold" target competitor cells. These findings indicate that the target antigen of the Tc was expressed on both autologous ILT cells and lymphoma cells, and it may be different from ATLV antigens detected by serologic methods. In addition, the data suggested allogeneic restriction of the Tc in that the preferentially killed allogeneic ATLV-bearing cells share several HLA antigens.
Regulation of the immune response to tumor antigens. X. Activation of third-order suppressor T cells that abrogate anti-tumor immune responses.
Authors: Schatten S , Drebin J A , Perry L L , Chung W , Greene M I
Published: December 2022

The experiments described further define the suppressor T cell pathway in the S1509a tumor system. We demonstrated previously that S1509a-induced Ts1, TsF1, and Ts2 specifically suppress in vivo Ly1+2- T cell-dependent responses to S1509a and that Ts1 suppress in vivo Ly-1+2- T cell-mediated proliferative responses to S1509a. We have now shown that in vivo administration of either S1509a-induced TsF1 or TsF2 suppresses both in vivo and in vitro Ly-1+2- T cell-mediated responses to S1509a. Furthermore, we revealed the existence of Ts3, which are activated by S1509a tumor antigen and TsF2, in this murine tumor system. Finally, we demonstrated that cyclophosphamide abrogates the suppressive effect of TsF2 but not that of Ts3. These results are discussed with respect to T cell-mediated suppression in other murine tumor systems and the possible pivotal role for a tumor antigen-presenting cell in activating Ts3 in the S1509a tumor system.
Human newborn autologous mixed lymphocyte response: frequency and phenotype of responders and xenoantigen specificity.
Authors: Hayward A R , Mori M
Published: December 2022

The response of human newborn lymphocytes in autologous mixed lymphocyte culture was examined for specificity (by restimulation), responder cell phenotype, and responder cell frequency. When the newborn T cells were separated from non-T cells by rosetting with sheep erythrocytes (E) in fetal calf serum (FCS), approximately 1:20,000 T cells proliferated. These responders had specificity for E + FCS, were T4+, and were self-restricted. Significant numbers of responder T cells were not found when newborn T and non-T cells were separated by nylon wool. Responses in parallel autologous cultures of adult T cells showed that 1) adults had a higher frequency than newborns of E + FCS specific responders, 2) evidence for self specificity was lacking in restimulated cultures, and 3) occasional responses to antigen on the surface of monocytes could not be excluded.
Interferon is the suppressor of hematopoiesis generated by stimulated lymphocytes in vitro.
Authors: Zoumbos N C , Djeu J Y , Young N S
Published: December 2022

Lymphocytes that inhibit hematopoiesis may have a pathogenic role in some forms of bone marrow failure, and lymphocyte-mediated suppression may also be important in the normal regulation of bone marrow function. We have investigated the mechanism of in vitro suppression of hematopoiesis by T cells by using the methylcellulose colony culture system. Total peripheral blood T cells and separated subpopulations of helper (OKT4+) and suppressor (OKT8+) cells that have been stimulated by exposure to lectin suppress autologous colony formation by bone marrow myeloid (CFU-C) and erythroid (BFU-E) progenitor cells. Medium conditioned by these cells is also inhibitory, indicating that the suppressor activity is a soluble factor. A strong correlation existed for the concentration of interferon and the degree of hematopoietic suppressor activity in these supernatants; both activities peaked at days 3 to 5 of incubation and had sharply declined by day 7. Interferon production was enhanced by exposure of lymphocytes to sheep red blood cells during the rosetting procedure. Specific antiserum and a monoclonal antibody directed against gamma-(immune) interferon abrogated the inhibitory activity for hematopoiesis produced by lectin-stimulated T cells; an antiserum to alpha-interferon was generally much less effective in neutralizing activity. We infer from these results that gamma-interferon is the mediator of hematopoietic suppression generated by lectin-treated T-cells.
A new lymphocyte surface antigen defined by a monoclonal antibody (9F3) to the T cell population expanding in MRL/Mp-lpr/lpr mice.
Authors: Dumont F J , Habbersett R C , Nichols E A
Published: December 2022

This report is a description of the pattern of reactivity of a rat monoclonal antibody (MAb 9F3) directed to the abnormal T cells expanding in MRL/Mp-lpr/lpr (MRL/lpr) mice. By using single- and two-color flow cytofluorometry analysis, this MAb was found to stain brightly 90 to 98% of lymph node (LN) T cells from MRL/lpr, C57BL/6-lpr/lpr (B6/lpr), and C3H/HeJ-lpr/lpr mice, and approximately 10 to 50 times less intensely 55 to 70% of T cells from control congenic +/+ mice. The study of in vitro proliferative responses of sorted cells from MRL/+ mice demonstrated that 9F3- T cells react better to phytohemagglutinin and allogeneic cells but less well to concanavalin A (Con A) than 9F3+ T cells. Upon Con A-induced blastogenesis, 65% of 9F3- T cells became 9F3+ whereas all 9F3+ remained 9F3+. Although only 15% of thymocytes, which included hydrocortisone-resistant population, were 9F3+ in +/+ mice, up to 60% of bright 9F3+ cells were detectable in the thymus of lpr-bearing mice after the onset of lymphoproliferation. Moreover, in both lpr-bearing and normal mice, the 9F3 MAb stained the totality of resting or mitogen-activated B cells. The 9F3 MAb also reacted with 100% of bone marrow (BM) cells, irrespective of the lpr gene. However, a subset of bright 9F3+, large BM cells was increased in frequency in lpr-bearing mice compared to congenic controls. Although expressed on macrophages, granulocytes, and erythrocytes, the 9F3 antigen was not in liver, kidney, and brain tissue of MRL/+ mice. Because the cell and tissue distributions of the 9F3 antigen do not correspond to any previously described murine antigen, this antigen may represent a new surface marker that should prove useful for studying lymphohemopoietic cell differentiation in normal and lpr-bearing mice.
Glycolipid antigens with blood group I and i specificities from human adult and umbilical cord erythrocytes.
Authors: Okada Y , Kannagi R , Levery S B , Hakomori S
Published: December 2022

Neutral glycolipids and gangliosides of umbilical cord and adult human erythrocytes were separated by high performance liquid chromatography, and each fraction was analyzed by direct binding of anti-I (Ma) and anti-i (Den) on solid phase glycolipid-lecithin-cholesterol. The I- and i-active glycolipids were isolated and their structures were determined by methylation analysis and direct probe mass spectrometry. The major I antigen in adult erythrocytes, showing a remarkable binding activity with anti-I(Ma), was found in one neutral glycolipid fraction, designated fraction y4, which was identified as a mixture of two glycolipids of a new type, lactoisooctaosylceramide and monofucosyllactoisooctaosylceramide (for structures, see Table I). In addition, two gangliosides displaying direct binding activity with anti-I(Ma) were identified as monosialoganglioside G8, as previously described and disialosyllactoisooctaosylceramide, which showed the same level of I activity as the y4 glycolipid. The same ganglioside was recently isolated and characterized by Kundu and co-workers. The major i-active glycolipid antigen in umbilical cord erythrocytes, showing a strong binding activity with anti-i(Den), was a neutral glycolipid, x4a, which was identified as lactonorhexaosylceramide. This glycolipid without fucosyl or sialosyl substitution has not been isolated previously and was present as an obvious normal component of umbilical cord erythrocytes, but an extremely minor component of adult erythrocytes. Sialosyllactonorhexaosylceramide (G6) was isolated and characterized as a second i antigen of umbilical cord erythrocytes, but showed a very weak binding activity with the anti-i antibody. Although these sialosyl derivatives displayed only weak activity, the chemical quantity of the sialosyl derivatives is significantly large in fetal erythrocytes; therefore, Ii activity of human erythrocytes, in general, must be significantly dependent on sialosyl derivatives in addition to unsubstituted structures.
Modification of cellular protein sulfhydryl groups by activated soluble immune response suppressor.
Authors: Aune T M
Published: December 2022

Soluble immune response suppressor (SIRS), a product of murine Ly-2+ T lymphocytes, is activated to SIRSox by H2O2 produced by macrophages: SIRSox directly inhibits cell division by normal and neoplastic cells and antibody secretion by B lymphocytes. To examine the mechanism of SIRSox-mediated inhibition, a variety of cellular functions were measured after treatment of cells with SIRSox. These included respiration, glucose transport, microtubule content, glutathione content, production of H2O2 or superoxide anion, and the activities of a variety of different enzymes. Several cellular activities or measurements were inhibited or lowered after SIRSox-treatment, including cell division, microtubule content, glutathione reductase activity, and thioredoxin reductase activity; inhibition was partially reversed by the sulfhydryl reducing agent dithiothreitol. Protein sulfhydryl content of P815 mastocytoma cells and several other cell types was lowered by 35 to 45% after exposure to SIRSox. Protein sulfhydryl loss was also partially restored after incubation with dithiothreitol. Sulfhydryl loss was not due to cell lysis. In addition, treatment of crude cellular particulate fractions with SIRSox resulted in protein sulfhydryl loss and formation of protein sulfenyl derivatives. A comparison of the amount of SIRS and H2O2 present to the number of protein sulfhydryls lost or sulfenyl derivatives formed suggests that SIRSox acts catalytically, serves as a co-factor in protein sulfhydryl oxidation, or that it activates a second pathway that is directly responsible for sulfhydryl oxidation.
Adenosine receptors on mouse bone marrow-derived mast cells: functional significance and regulation by aminophylline.
Authors: Marquardt D L , Walker L L , Wasserman S I
Published: December 2022

Adenosine receptors on mouse bone marrow-derived mast cells were identified by functional criteria and radioligand binding. The stimulated release of beta-hexosaminidase from these cells was significantly augmented by the simultaneous addition of secretagogue and adenosine, NECA, or L-PIA. Similar enhancement of pre-formed mediator release occurred after a 10-min preincubation with adenosine. Resting mast cell cAMP levels increased within 15 sec after the addition of adenosine, and remained elevated for at least 60 sec. Although the antigen-or A23187-induced release of beta-hexosaminidase was markedly potentiated by exogenous adenosine, the stimulated release of [14C]-labeled arachidonic acid metabolites was minimally affected by adenosine, suggesting a differential effect of adenosine on granule-associated release as compared to generated mediator release. Bone marrow mast cells exhibited 5470 +/- 740 [3H]adenosine binding sites/cell, with a binding affinity of 24.4 +/- 3.8 nM. Cells cultured in the presence of 100 microM aminophylline for 6 days were hyperresponsive to exogenous adenosine, releasing a maximum of 162% of the amount of beta-hexosaminidase released from control cells in the presence of adenosine. The number of [3H]adenosine binding sites on the xanthine-treated cells increased to 156% of control values, suggesting an up-regulation of adenosine receptors induced by chronic exposure to an adenosine receptor antagonist. Mouse bone marrow mast cells possess functionally significant adenosine receptors that are regulated by aminophylline and that, when stimulated, produce many alterations in the mast cell secretory process.
The synergistic anti-proliferative effect of gamma-interferon and human lymphotoxin.
Authors: Lee S H , Aggarwal B B , Rinderknecht E , Assisi F , Chiu H
Published: December 2022

Highly purified preparations of rHuIFN-gamma and HLT synergistically inhibit the in vitro proliferation of WI38VA13 cells, a SV40 virus-transformed derivative of the human cell line W138. The growth of nontumorigenic WI38 cells, established from normal diploid embryonic lung tissue, was not inhibited under identical incubation conditions; rMuIFN-gamma and HLT have the same in vitro effects on the murine melanoma B16. Thus, the combination of rMuIFN-gamma and HLT synergistically inhibits the growth of B16 cells, although incubation with either lymphokine alone has no significant anti-proliferative effect. Flow cytometric analysis of the cell cycle of asynchronous cells indicates that when B16 melanoma cells are treated with rMuIFN-gamma and HLT in combination, more cells accumulate in the G0/G1 phase than when the cells are treated with rMuIFN-gamma alone. It appears that populations of rapidly proliferating tumor cells, which normally contain a large percentage in the S+G2+M phases of the cell cycle, become arrested in the quiescent G0/G1 phase by the combination of IFN-gamma and HLT, resulting in the inhibition of cell growth.
Inhibition of human B cell activation by diterpine forskolin: interference with B cell growth factor-induced G1 to S transition of the B cell cycle.
Authors: Muraguchi A , Miyazaki K , Kehrl J H , Fauci A S
Published: December 2022

We have previously demonstrated that a series of sequential steps are involved in the induction of a resting human B cell to proliferate. In this system, initial activation signals were delivered by either in vivo or in vitro stimulation, and proliferative signals were delivered by a B cell growth factor (BCGF) that was derived from a human T-T cell hybridoma. The involvement of adenosine 3',5'-monophosphate (cAMP) in regulating the growth and differentiation of lymphocytes has been of considerable interest. Diterpine forskolin has been reported to be a unique adenylate cyclase activator in membranes from mammalian tissues that results in elevations of intracytoplasmic cAMP. We have examined the effect of this drug on the progression of human B cells through their activation cycle. It was found that forskolin causes a rapid and sustained increase of cytoplasmic cAMP in purified small and large B cells. In the in vitro costimulation of small resting B cells with anti-mu plus BCGF, forskolin inhibited the proliferative response of B cells in a dose-dependent manner. This forskolin-mediated suppression of B cell proliferation was found when the drug was added to the cultures as late as 36 hr after initial stimulation. Of note is the fact that the anti-mu-induced RNA synthesis as well as cell enlargement in resting B cells was not inhibited by the addition of forskolin, whereas BCGF-induced proliferative response of activated B cells was markedly inhibited by the drug. Thus, these data demonstrate that forskolin, an agent that elevates cytoplasmic cAMP levels, has relatively little effect on early events in the human B cell cycle (G0 to G1) transition, but selectively inhibits the progression of BCGF-induced G1 to S phase transition of B cells.
Thymocyte differentiation activity from the cloned monocyte/macrophage cell line RAW 264.7. Alterations in the expression of immature thymocyte surface antigens.
Authors: McKernan L N , Largen M T
Published: December 2022

The cloned monocyte/macrophage cell line RAW 264.7 was previously shown to produce thymocyte mitogenic and co-mitogenic activity that eluted from a Sephadex G-75 column not only at approximately 16,000 daltons, the m.w. described for interleukin 1 (IL 1), but also at 30,000 to 40,000 daltons. The studies reported here indicate that the 30,000 to 40,000 dalton molecule has thymic differentiating activity. Thymocytes from A/J mice were fractionated on discontinuous BSA gradients, which yielded populations of cells enriched for immature and mature cells. The cells found at the interface between 35 and 29% BSA (band 1 cells), which are the most immature, were cultured for 48 hr with highly purified IL 1, with the 30,000 to 40,000 dalton form of thymocyte co-mitogenic activity obtained after Sephadex G-75 chromatography and chromatofocusing chromatography, or with media alone. The surface antigens TL-3, H-2Kk, Thy-1.2, Lyt-1, and Lyt-2 were examined by immunofluorescence. It was found that the highly purified 30,000 to 40,000 dalton species of co-mitogenic activity induced a significant increase in the content of surface H-2Kk, a decrease in TL-3, and a very small decrease in Thy-1.2 on the cell surface, whereas IL 1 was not capable of inducing a change in these surface antigens. There was no change in Lyt-1 on the surface of band 1 thymocytes after incubation with either IL 1 or the 30,000 to 40,000 dalton species. The 30,000 to 40,000 dalton species caused a significant decrease in the percentage of cells staining positive for Lyt-2, whereas IL 1 caused a smaller but significant decrease in Lyt-2. These changes in the surface markers TL-3, H-2Kk, and Thy-1.2 are consistent with changes that occur during thymocyte differentiation. It was also observed that the proliferative response to the 30,000 to 40,000 dalton form and IL 1 increased with increasing functional maturity of each band of thymocytes when used in the thymocyte mitogenic assay. However, only the 30,000 to 40,000 dalton form was capable of inducing a proliferative response in the immature band 1 thymocytes in the thymocyte co-mitogenic assay. These results indicate that the RAW 264.7 cells produce a factor that has, in addition to thymocyte co-mitogenic activity, thymocyte differentiation activity, and this factor is distinct from IL 1.
Evidence for restriction of the ability of complement to lyse homologous erythrocytes.
Authors: Houle J J , Hoffmann E M
Published: December 2022

This research explored the possibility that a mechanism for the inhibition of C exists that is capable of restricting lysis of antibody-sensitized homologous E. The ability of C to lyse E from six species was compared by using a passive lysis system that employed E coated with B. abortus LPS and bovine antibodies specific for B. abortus. A system was developed that permitted comparisons of the hemolytic efficiencies of heterologous and homologous C target cell combinations. C from all of the species tested lysed heterologous E effectively, but homologous E were poorly lysed. Furthermore, a serum factor appeared to be involved with restriction of lysis of homologous E:LPS:Ab by heterologous C, and the restriction was specific for homologous E. Human E:LPS:Ab that were washed after incubation with CH were resistant to subsequent lysis by heterologous C. This restriction did not occur at 0 degrees C or in the absence of antibodies. Incubation of homologous C with EH:LPS:Ab resulted in consumption of C1, C2, C3, and C4, but not C5. The treated CH would lyse sheep EAC1423, but it would not lyse sheep EA, EACI, or EAC142. Late reacting C components (C5 through C9) were not detectable on EH:LPS:Ab after incubation with CH, but C4 and C3 were bound to the cells. Rat late-reacting C components (rat C-EDTA) were capable of detecting C3 convertase sites on EH:LPS:Ab that had been reacted with CH. However, homologous late-reacting components and guinea pig late-reacting components were unable to detect the C3 convertase sites.
The fate of IgE bound to rat basophilic leukemia cells. III. Relationship between antigen-induced endocytosis and serotonin release.
Authors: Furuichi K , Rivera J , Isersky C
Published: December 2022

We have previously shown that, unlike monomeric IgE, chemically derived dimers, trimers, and heavier oligomers of IgE were internalized efficiently. This finding suggested that endocytosis, like mediator release, is triggered by cross-linking of the cell surface receptors for IgE. In the present study, we analyzed the temporal and functional relationships between the two events. We used rat basophilic leukemia cells (RBL-HR+-2H3) and rat peritoneal mast cells, which were allowed to bind monomeric 125I mouse IgE hybridoma anti-dinitrophenyl (HI-DNP-E-26-82), and the polyvalent antigen 131I-dinitrophenylated human serum albumin (DNP15-HSA). We found that at 37 degrees C, 50% of the cell surface-bound immune complexes were internalized rapidly (t1/2 3 to 5 min) by RBL-HR+-2H3 cells with only minimal reduction (1/3) in the extent of internalization when very few of the receptors (approximately 5%) were saturated with IgE. Normal mast cells internalized cell surface-bound immune complexes at a similar rate (t1/2 4 to 5 min). Unlike serotonin release, internalization was independent of extracellular calcium and continued to increase as the ratio of DNP15-HSA to IgE increased 10- to 100-fold over the ratio required for optimal histamine release. In the RBL cells, internalization preceded serotonin release, reaching a peak at about 10 min, while the release (t1/2 13 to 19 min) continued for up to 60 min. Presumably, some of the cross-linked IgE internalized less effectively and continued to trigger serotonin release. The reverse relationship between the rates of internalization and release (t1/2 less than 1 min) was found in normal rat mast cells. We conclude that although cross-linking of two or more receptors triggered both endocytosis and exocytosis, the two events are not necessarily sequential.
Induction of crisis forms in the human malaria parasite Plasmodium falciparum by gamma-interferon-activated, monocyte-derived macrophages.
Authors: Ockenhouse C F , Schulman S , Shear H L
Published: December 2022

Previous investigations on the mechanism by which the host mounts an immune response against the human malaria parasite Plasmodium falciparum have not resolved whether cell-mediated responses, in the absence of circulating anti-Plasmodial antibodies, can effect the destruction of the intraerythrocytic parasite. We report that the intraerythrocytic parasite P. falciparum is lethally susceptible to the imposition of oxygen-dependent and oxygen-independent factor(s) released by interferon-gamma-activated, monocyte-derived human macrophages. In addition, trophozoite-schizont stage intraerythrocytic parasites were killed on exposure to small amounts of H2O2 generated in cell-free enzyme assays. Although parasiticidal activity was markedly enhanced by the addition of lactoperoxidase and KI, killing was abrogated by the addition of catalase. The ability of freshly isolated human monocytes, monocyte-derived macrophages (MDM), and lymphokine-activated MDM to kill or inhibit the growth and multiplication of the malaria parasites was assessed. Parasites were killed when exposed to monocytes or lymphokine-activated MDM, but not when exposed to nonactivated macrophages. The capacity to activate MDM for microbicidal activity was abrogated on neutralization of crude lymphokines or recombinant interferon-gamma with a monoclonal antibody prepared against interferon-gamma. The intraerythrocytic parasites surviving the cytotoxicity assay were inhibited in their development and appeared to be degenerating, a characteristic of "crisis" forms. Killing of P. falciparum correlated positively with the magnitude of the oxidative response, as evidenced by the reduction of nitroblue tetrazolium to formazan in the mononuclear phagocytes, and by the detection of secreted H2O2. Of particular interest was the observation that only the later developing stage of the intracellular parasite triggered the respiratory burst in the absence of antibody. A role for oxygen-independent parasiticidal factors was suggested by the finding that lymphokine-activated macrophages from a patient with chronic granulomatous disease were able to partially inhibit the growth of P. falciparum, although oxidative metabolism in these cells was impaired.
Genetically determined resistance to mouse hepatitis virus 3 is expressed in hematopoietic donor cells in radiation chimeras.
Authors: Dupuy J M , Dupuy C , Décarie D
Published: December 2022

Differences in mouse hepatitis virus 3 (MHV3) sensitivity among mouse strains are mainly determined by H-2-related and -nonrelated genetic factors. Reciprocal chimerism was therefore established between two H-2a compatible pairs of strains that differ widely in their susceptibility to MHV3: a) A/J and B10.A, respectively resistant and highly susceptible; b) A/J and A/Sn, respectively resistant and semisusceptible. Chimeric mice were challenged with 100 LD50 of MHV3, 30 or 90 days after X-irradiation (900 R) and bone marrow reconstitution. Results showed that sensitivity of recipients was similar either to that of the recipient strain or to that of the donor strain when chimeric mice were tested 30 or 90 days, respectively, after reconstitution. In addition, no paralysis occurred in surviving animals. These data indicate, therefore, that resistance or susceptibility to MHV3 is expressed intrinsically in some population(s) of hematopoietic-derived cells, which is radioresistant and has a life span of more than 30 days and less than 90 days. Additional experiments showed that X-irradiated A/J recipients reconstituted with A/J bone-marrow cells were protected against MHV3 challenge with spleen cells, with a mixture of spleen cell populations or of adherent spleen cells and thymocytes originating from A/J donors. Transfer of protection to recipients by using similar cell populations provided by semisusceptible A/Sn donors required the administration of five times more cells. Results suggest that two complementary mechanisms are required to confer resistance to MHV3: a) a gene(s) for resistance that may operate at the level of macrophages, and b) cells capable of mounting an efficient immune response. The reduced efficiency of A/Sn spleen cells suggests that semisusceptibility to MHV3 may be related to partial quantitative or functional immune defect.
An antigen characteristic of hairy cell leukemia cells is expressed on certain activated B cells.
Authors: Posnett D N , Wang C Y , Chiorazzi N , Crow M K , Kunkel H G
Published: December 2022

A murine monoclonal antibody, anti-HC2, which reacts with hairy cell leukemia cells, was used to define the normal cell equivalent for the leukemic hairy cell. This antibody stained the membranes of 2.24% of normal peripheral blood lymphocytes. These cells were more common in T cell-depleted cell populations, and bore membrane immunoglobulin. Some B lymphoblastoid cell lines were also anti-HC2-positive. Examination of normal B cell subpopulations revealed that HC2-positive cells frequently co-express an activation antigen and membrane IgG. Cell populations enriched for HC2-positive cells contained the bulk of PWM-responsive B cells. These data suggest that HC2-bearing B cells are activated B cells. B cell differentiation induced in vitro by PWM with T cell helper factors resulted in an increase in the number of HC2-positive cells at day 4 or 5 of culture. The HC2-positive cells were no longer present at the time of maximal plasma cell differentiation on day 7 of culture. By using B cells from a patient with the hyper-IgM syndrome that are incapable of immunoglobulin heavy chain class switching, it was demonstrated that HC2 expression did not require prior heavy chain switch. The m.w. of the HC2 antigen was 52 to 63 KD. Four bands with different isoelectric points were discerned on two-dimensional analysis. We suggest that hairy cell leukemia represents a malignancy of activated B cells. A unique stage of B cell differentiation is identified by the HC2 activation antigen.
Generation of heavy chain-loss mutants in a B cell hybrid mediated by syngeneic idiotype-specific spleen cells.
Authors: Ghosh S K , Bankert R B
Published: December 2022

Idiotype-specific spleen cells from appropriately primed BALB/c mice cause a marked and irreversible suppression of the membrane and secreted forms of idiotype-positive immunoglobulin (Ig) of an antigen-specific B cell hybrid clone (2C3E1). The suppression of this BALB/c B cell line has been observed in vitro and in vivo, and appears to require intimate contact between effector spleen cells and target 2C3E1 cells. The observed suppression in the 2C3E1 cell line is due to an induced mutation or a selection of pre-existing mutants within the 2C3E1 cell population, because the resultant light and heavy chain-loss variants are phenotypically stable in vitro and in vivo in the absence of any further active suppression. Biochemical analysis of the 2C3E1 cells after this suppression indicates that all of the variants are negative for the production of idiotype-positive Ig. Heavy chain synthesis by the variants is almost totally eliminated, and light chain synthesis is decreased by 10 to 90%. Spleen cells from identically primed nude mice do not induce any alteration in the 2C3E1 cell line, suggesting that induction or selection of the heavy and light chain-loss mutants requires the presence of mature T lymphocytes. The generation of idiotype-negative 2C3E1 variants during the period of tumor growth in the spleen (but not elsewhere) may represent one mechanism by which this tumor escapes the host's immune recognition.
An association between susceptibility to experimental autoimmune uveitis and choroidal mast cell numbers.
Authors: Mochizuki M , Kuwabara T , Chan C C , Nussenblatt R B , Metcalfe D D , Gery I
Published: December 2022

An association was found in rats of different strains between the susceptibility to EAU and the number of mast cells in the choroid of the eye. High responder rats (Lewis, CAR) had strikingly more choroidal mast cells than the low responder BN animals, whereas intermediate numbers of mast cells were found in the F1 hybrids of Lewis and BN (LBNF), which exhibited an average level of susceptibility to EAU. LeR rats, derived from the Lewis strain, developed EAU only when treated with B. pertussis, and their number of choroidal mast cells was only about 1/5 of that found in the Lewis rats. Unlike the differences in the number of choroidal mast cells, small variations were found in the skin mast cell numbers of the tested rats. It is proposed that the number of local mast cells may be one mechanism by which the susceptibility to an organ-specific autoimmune disease is genetically regulated.
Studies on the mechanism of the enhancement of delayed-type hypersensitivity by pertussigen.
Authors: Sewell W A , Munoz J J , Scollay R , Vadas M A
Published: December 2022

The potentiation of delayed-type hypersensitivity (DTH) reactions by pertussigen, a protein toxin from Bordetella pertussis, has been studied in adoptive transfer assays. Lymph node or spleen cells from mice treated with or without pertussigen at the time of immunization with protein antigens were transferred to naive, syngeneic recipients that were challenged with antigen. Cells from donors treated with pertussigen had the capacity to transfer vigorous, antigen-specific DTH reactions. Cells from immunized donors not given pertussigen transferred little or no DTH. These results indicate that pertussigen is able to augment DTH reactions by potentiating the antigen reactivity of cell populations in lymphoid organs. The phenotype of the effector cells induced by pertussigen was Thy-1 positive, L3T4 positive, and Ly-2 negative. Cells from mice given pertussigen and an irrelevant antigen had no influence on specific DTH responses, suggesting that pertussigen enhances the activity of the antigen-specific cell type mediating DTH. The effect of pertussigen and of immunization on the lymphocyte subpopulations present in the lymph nodes was studied by analysis of suspensions of lymph node cells by flow cytometry. In immunized and in nonimmune mice, pertussigen increased the ratio of Ly-2-negative:Ly-2-positive T cells, and reduced the overall proportion of B cells. In immunized mice, pertussigen induced a much higher proportion of large dividing cells from 5 days after sensitization onwards. The relevance of these changes in lymphocyte behavior to the development of enhanced and prolonged DTH in mice given pertussigen is discussed.
Regulation of immune responses by I-J gene products. IV. Distinct suppressor factors derived from "nonsuppressor" A strain mice.
Authors: Lei H Y , Waltenbaugh C
Published: December 2022

A/J, A/WySn (H-2a, I-Jk), and B6 (H-2b, I-Jb) mice are poly(Glu50Tyr50)(GT) nonresponders. GT preimmunization does not suppress humoral immune responses to the immunogenic form of GT, GT-MBSA (GT-methylated bovine serum albumin), in these strains. B6 mice produce an idiotypic GT-specific T cell-derived suppressor factor (GT-TsF1) after GT injection, but cannot be suppressed by this or any other GT-TsF1. H-2a mice do not produce GT-TsF1, but can be suppressed by B6 GT-TsF1. Injection of cellfree B6 GT-TsF1 into H-2a mice leads to the production of two distinct I-Jk GT-TsF. We have termed these GT-TsF2 and GT-TsF3 on the basis of their kinetic activity and antigenic markers. Anti-idiotypic I-Jk GT-TsF2 is suppressive at a time intermediate between GT-TsF1 and GT-TsF3. Idiotypic I-Jk GT-TsF3 suppresses GT-MBSA PFC responses the day of PFC assay. GT-TsF3, not GT-TsF1, suppresses B6 GT-MBSA PFC responses, indicating that these idiotypic factors are distinct. Our results establish that H-2a lack or have a defective Ts1 cell for GT suppression, and H-2b mice lack a functional Ts2 cell. These findings offer strategies for circumventing suppressor T cell defects in "nonsuppressors."
Phorbol esters inhibit murine B cell differentiation to immunoglobulin secretion but not proliferation.
Authors: Isakson P C , Simpson L
Published: December 2022

Bacterial lipopolysaccharide (LPS) induces resting B cells to proliferate and to secrete IgM. We have found that addition of phorbol esters (PE) such as phorbol myristate acetate (PMA) to murine B cells specifically inhibits LPS-induced IgM secretion but not proliferation. PMA is extremely potent, with half-maximal inhibition occurring at about 10 pM. The effect on B cells appears to be due to interaction with PE receptors, because a series of PE have similar potencies for tumor promotion, binding to receptors, and inhibition of IgM secretion. PE also inhibit IgM secretion induced by T cell-derived lymphokines and LPS-induced IgG secretion. Results of these studies suggest that the protein kinase C, with which PE interact, plays an important role in the regulation of B cell differentiation and may also provide a powerful tool for dissecting molecular events involved in induction of Ig secretion.
T cell regulation of myelopoiesis: analysis at a clonal level.
Authors: Griffin J D , Meuer S C , Schlossman S F , Reinherz E L
Published: December 2022

Colony-stimulating factor (CSF) production by a series of cloned human T lymphocyte cell lines was examined by substituting cloned T cells for peripheral blood mononuclear cells in the feeder layer of a double-layer agar CFU-C assay system. Of 12 T cell lines tested, all produced CSF when stimulated by specific antigen, whereas CSF production in the absence of stimulation was generally negligible. In the case of soluble antigen-specific (ragweed or tetanus toxoid) clones, this required both nominal antigen and the appropriate MHC gene product on autologous antigen-presenting cells, whereas in the case of clones specific for EBV-transformed B cell lines (allogeneic or autologous), surface-bound EBV-related antigen and MHC was necessary. When tested in this manner, CSF production by different cloned T cells was heterogeneous in both amount and subclass. Thus, although most clones stimulated growth of granulocyte, monocyte, and eosinophil colonies, certain clones were identified which preferentially stimulated some colony types but not others. This heterogeneity was particularly evident with respect to eosinophil colony production. In addition, a soluble inhibitor of granulocyte colony growth was produced by one clone. These findings provide further support for the notion that antigen-specific T cells may, on activation, regulate myelopoiesis in a precise way, and provide a possible cellular basis for selective eosinophilia, monocytosis, or neutrophilia seen in certain disease states.
Autoimmune effector cells. VI. Transfer of experimental allergic encephalomyelitis with spleen cells activated in mixed lymphocyte cultures.
Authors: Hayosh N S , Simon L L , Swanborg R H
Published: December 2022

Although spleen cells (SpC) from Lewis rats that have been immunized with guinea pig myelin basic protein in complete Freund's adjuvant do not transfer experimental autoimmune encephalomyelitis (EAE) to syngeneic recipients, we report that effector cells of EAE can be activated in SpC suspensions during mixed lymphocyte reactions (MLR) to allogeneic SpC. Effector cell activation correlates with interleukin 2 (IL 2) production. The results are consistent with the hypothesis that autoreactive cells may be generated as a result of an immune response to exogenous antigens.
Effects of monoclonal antibodies directed against murine T lymphocyte cell surface antigens on lymphokine production by cloned T lymphocytes reactive with class I MHC or Mls alloantigens.
Authors: Lancki D W , Prystowsky M B , Vogel S N , Beller D I , Dialynas D P , Fitch F W
Published: December 2022

Monoclonal antibodies recognizing murine T lymphocyte cell surface structures implicated in T lymphocyte-mediated cytolysis, including Lyt-2, L3T4, LFA-1, and a cytolytic T lymphocyte (CTL) clonotypic determinant, were used as probes to investigate the role of these structures in lymphokine production by T cell clones induced by antigen or lectin. The clone-specific antibody 384.5 bound to and inhibited antigen-induced lymphokine production by the L3 CTL clone, but did not affect lymphokine production by other T cell clones. Antibodies against the T cell surface structures Lyt-2 or L3T4, which are expressed by mutually exclusive T cell subsets, inhibited antigen-induced lymphokine production by class I major histocompatibility complex (MHC) antigen-reactive CTL clones or an M1s-reactive helper T lymphocyte (HTL) clone, respectively. Antibody against the broadly distributed LFA-1 molecule inhibited antigen-induced lymphokine production by all of the clones tested. Lectin-induced lymphokine production by cloned T cells was not inhibited by the clonotypic antibody, anti-Lyt-2, or anti-LFA-1; slight inhibition of the HTL clone was observed with the anti-L3T4 antibody. None of these structures appear to be uniquely involved with a particular functional response. Our results suggest that each of these structures is involved with the interactions between the effector cell and the stimulating cell leading to lymphokine production.
Stimulation of basophil and rat mast cell histamine release by eosinophil granule-derived cationic proteins.
Authors: Zheutlin L M , Ackerman S J , Gleich G J , Thomas L L
Published: December 2022

Major basic protein (MBP), an arginine-rich basic polypeptide that constitutes the crystalloid core of the large specific eosinophil granule, has previously been shown to stimulate noncytolytic histamine release from human basophils and rat mast cells by an IgE-independent mechanism. Two additional basic polypeptides present in eosinophil granules, eosinophil cationic protein (ECP) and eosinophil-derived neurotoxin (EDN), were examined for similar activity in the present study. Acid-solubilized eosinophil granules were fractionated by chromatography on a Sephadex G-50 column. Incubation of basophil-containing human mononuclear cells with the individual column fractions demonstrated that histamine release occurred only with the fractions that contained MBP. The selectivity of the basophil response for MBP was confirmed by using equimolar concentrations of purified MBP, ECP, and EDN. In contrast, both MBP and ECP, but not EDN, stimulated histamine release from purified rat peritoneal mast cells. Reduction and alkylation of the MBP molecule diminished the response of human basophils to MBP but enhanced the potency of the molecule with rat mast cells. The distinct potency of MBP as a stimulus for histamine secretion from human basophils suggests that eosinophil release of MBP may be a specific event in the augmentation of immediate hypersensitivity reactions and other disorders characterized by eosinophilia.
Human natural killer cell recognition of herpes simplex virus type 1 glycoproteins: specificity analysis with the use of monoclonal antibodies and antigenic variants.
Authors: Bishop G A , Marlin S D , Schwartz S A , Glorioso J C
Published: December 2022

The specificity of human natural killer (NK) cells for herpes simplex virus type 1 (HSV-1)-infected WISH cells was examined by using virus-specific antibodies to block NK killing and by using targets infected with virus variants that express antigenically altered cell surface glycoproteins. Fab fragments of human immunoglobulin (Ig) derived from an HSV-1 seropositive individual blocked NK activity against HSV-1-infected cells, whereas Fab fragments from a seronegative individual had no effect. Fab fragments of monoclonal antibodies specific for different epitopes of the HSV-1 glycoproteins gB and gC blocked NK activity, suggesting that NK cells interact directly with viral glycoproteins. In contrast, a control (HSV-unreactive) monoclonal antibody did not block NK activity. Further evidence that the blocking activity was associated with HSV glycoproteins was obtained by showing that: a) gC-specific antibodies did not decrease NK activity against cells infected with an HSV-1 mutant which does not insert gC into infected cell surface membranes; b) gB-specific antibodies did not reduce NK cytotoxicity against target cells infected with HSV-1 in the presence of 2-deoxy-D-glucose (2dG), which prevents cell surface expression of gB; c) HSV-1 glycoprotein-specific antibodies did not alter NK activity against uninfected cells or against K562, an NK-susceptible tumor cell line; and d) a monoclonal antibody reactive with a cell membrane glycolipid markedly reduced NK reactivity against K562 but had little effect on NK reactivity against HSV-1-infected cells, suggesting that NK cells recognize different determinants on the two types of target cells. Analysis of the fine specificity of NK cell recognition of HSV-1 glycoproteins was performed by using target cells infected with antigenic variants of HSV-1 referred to as monoclonal antibody-resistant (mar) mutants. mar mutants are altered in distinct epitopes of gB or gC, resulting in loss of recognition by the selecting virus-neutralizing monoclonal antibody. NK activity was reduced against cells infected with some, but not all, of these variants. Together, these findings demonstrated a positive correlation between expression of HSV-1 glycoproteins gB and gC and susceptibility of HSV-1-infected target cells to NK activity, implicating the viral glycoproteins as cell surface components recognized by NK cells. Moreover, mutations affecting glycoprotein epitopes defined by monoclonal antibodies also influenced the interaction between NK effector cells and HSV-1-infected target cells, suggesting that viral antigenic sites recognized by NK cells overlap with those recognized by anti-HSV-1 antibodies.
Roles of V antigen in promoting virulence and immunity in yersiniae.
Authors: Une T , Brubaker R R
Published: December 2022

It is established that yersiniae harboring an approximately 45-megadalton Vwa-plasmid can produce V and W antigens (Vwa+), and that sera containing anti-V provides passive protection to mice against Yersinia pestis. This observation was extended by the use of monospecific anti-V prepared by injecting rabbits with partially purified V, absorption of antisera with a Vwa- extract, and then separation of gamma-globulin by traditional processes of fractionation or by affinity chromatography. These preparations provided passive protection against 10 minimum lethal doses of virulent Y. pestis KIM, Yersinia pseudotuberculosis PB1, and Yersinia enterocolitica WA. Kinetics of elimination of these Vwa+ yersiniae from organs and blood of passively immunized mice closely resembled those of avirulent Vwa- mutants from normal mice. Injection into mice of sterile crude extracts of Y. pseudotuberculosis PB1 containing V promoted significant survival and retention of Vwa- mutants of Y. pestis, Y. pseudotuberculosis, and Y. enterocolitica. This effect was eliminated by the removal of V before injection by precipitation with monospecific antibody. These results indicate that V antigen per se is the major virulence factor mediated by Vwa-plasmids.
Alloimmune memory is absent in Hymeniacidon sinapium, a marine sponge.
Authors: Smith L C , Hildemann W H
Published: December 2022

The southern California sponge Hymeniacidon sinapium can recognize and reject transplanted allogeneic tissues. This species reveals a complex rejection response that involves cellular infiltration into the graft interface and a deposition at that site of a fibrous wall. The response then proceeds to a cytotoxic reaction in which the cells in the graft zone become necrotic and slough off, resulting in allogeneic tissue separation. The rate and intensity of this response varies with the genetic constitutions of the paired sponges and the temperature of the water. In experiments employing pre-sensitized second-set and unrelated third-party graftings, with changes in temperature, sensitization times, intervals before regrafting, and experimental sites, this species did not exhibit alloimmune memory (i.e., accelerated second-set rejection). When the allorejection described here for H. sinapium is taken in conjunction with rejection responses reported for other sponge species, it is apparent that sponges have two major methods for responding to allogeneic contacts: barrier formation or cytotoxicity. The rejection method seems to correspond with the presence or absence of immune memory, and may, in part, be correlated with habitat and frequency of contacts with "non-self." The development of the rudimentary immune system in metazoans is discussed in relation to the rejection of naturally transplanted tissues in addition to challenges from pathogens.