bluuebunny/crossref_metadata_2025 huggingface dataset:
Influence of Y and X chromosomes on B cell responses in autoimmune prone mice.
Authors: Rosenberg Y J , Steinberg A D
Published: December 2022

BXSB mice, unlike other autoimmune strains, carry a Y chromosome-linked gene for accelerated autoimmunity, reflected by early onset disease in male but not female animals. To investigate whether the Y chromosome factor acts through other autosomal genes or is in itself sufficient for early disease, partially inbred congenic NZB.BXSB and NZW.BXSB mice were developed and studied for levels of Ig-secreting cells (IgFC) as a measure of their autoimmune state. In such mice, the BXSB contributed only the male chromosome and 3% of the autosomal genes. The results indicate that for full expression of autoimmunity, both the Y chromosome and one or more BXSB autosomal genes are required. In addition, the presence of the CBA/N xid gene results in a total reversal of any Y-linked effects.
Natural release of covalently bound C3b from cell surfaces and the study of this phenomenon in the fluid-phase system.
Authors: Venkatesh Y P , Minich T M , Law S K , Levine R P
Published: December 2022

Covalently bound C3b is released from cell surfaces (EAC1423 and zymosan-C3b) on incubation under physiologic conditions. The release of C3b from cell surfaces occurs by the cleavage of the covalent bond. Sodium dodecyl sulfate (SDS) abolishes the release, thereby indicating the requirement of the native structure of C3b in this process. The phenomenon of release of C3b from cell surfaces has also been observed in the fluid-phase system by using C3b-[3H]glycerol. The kinetics of the release of [3H]glycerol from C3b-[3H]glycerol were studied at 37 degrees C in 0.15 M phosphate buffer, pH 7.4. The first-order rate constant was found to be 0.028 +/- 0.003 hr-1. The release does not take place in either 8 M urea or 6 M guanidine hydrochloride, at pH 7.4. Under alkaline conditions, the rate of release is unaffected in the presence of SDS, indicating that the release in this pH range is not dependent on the native structure of the protein. From the Arrhenius plot in the temperature range 18 to 37 degrees C, an apparent activation energy for the hydrolysis reaction of 21.2 kcal/mol was calculated. The release phenomenon is exclusive for ester-linked complexes, as inferred by the absence of release of [3H]threonine from C3b-[3H]threonine, wherein the linkage is of the amide type. The presence or absence of the C3a portion of the molecule has no effect on the rate of release. The modification of the -SH group of C3i-/C3b-[3H]glycerol alters the rate of hydrolysis of the ester bond between C3i/C3b and [3H]glycerol. Protease inhibitors (PMSF, benzamidine HCl, and DFP) do not alter the rate of release, indicating that the hydrolysis reaction is not due to trace amounts of contaminating proteases. Thus, it appears that some chemical group(s) of C3i/C3b is (are) involved in the intramolecular hydrolysis of the ester bond between C3i/C3b and small molecules. This phenomenon may play an important role in the release of C3b from receptive surfaces once the biologic functions that require covalently bound C3b have been mediated.
Development of respiratory mucosal tolerance during Haemophilus influenzae type B infection in infancy.
Authors: Rosales S V , Lascolea L J , Ogra P L
Published: December 2022

Groups of patients with different forms of infection with Haemophilus influenzae type B (Hib), namely meningitis, epiglottitis, arthritis, and periorbital cellulitis, were evaluated for the appearance of serum IgG, IgA, IgM, and nasopharyngeal secretory (NPS) IgA (SIgA) antibody response to Hib capsular antigen at various intervals after the onset of clinical illness, by using an indirect enzyme-linked immunosorbent assay. The serum immune response was characterized by its predictable absence in infants under 23 mo of age, and in those with meningitis who, regardless of age, had high levels of circulating antigen. On the other hand, antibody response was frequently detected in the serum of older infants. Significantly, however, the appearance of SIgA antibody was demonstrated in virtually all patients with Hib infections under 23 mo of age. In addition, a positive correlation was observed between the concentration of antigen NPS, the level of SIgA activity in the NPS, and the absence of antibody response in the serum. These observations are strikingly similar to the development of systemic hyporesponsiveness (oral tolerance) observed after oral administration of certain infectious or nonreplicating antigens in experimental animals. It is suggested that similar mechanisms may underlie the immunologic abnormalities observed in the serum antibody response in infants with Hib meningitis.
Characterization of T lymphocyte colony-forming cells in the mouse. The colony-forming cell is confined to Lyt-1,2,3+ cell subsets in the thymus and the lymph nodes.
Authors: Röpke C
Published: December 2022

When cell populations from the thymus were studied with FACS, it was found consistently that the brightly labeled Thy-1.2+ populations contained very few T colony-forming cells (CFC), while these latter cells were numerous in the cell populations showing lower Thy-1.2 antigen density. This was paralleled by findings after peanut agglutinin (PNA) separation that showed enrichment of CFC in the PNA-negative medullary population, and by sorting based on TL, T-200, and H-2 determinants or light scatter properties of the cells. By FACS sorting of Lyt-labeled thymic cells, it was also shown that CFC were predominantly present in cell populations that were brightly Lyt-1+, and exclusively in populations that were Lyt-2+ and Lyt-3+. After FACS sorting of lymph node cells, no major differences in colony formation were found between dully- and brightly-labeled Thy-1.2+ or Lyt-1+ populations, or between lymphoid cells showing different light scatter characteristics. In addition, it was shown that CFC--like thymic CFC--were of the Lyt-1,2,3+ phenotype. It is concluded that the CFC may be present in several differentiation steps of Lyt-1,2,3+ cell lines, and that the frequency of these cells increases from the thymic cortex via the medulla and to peripheral lymphoid tissues.
The immune response to bacterial dextrans. IV. Antibody idiotypes.
Authors: Ivars F , Holmberg D , Cazenave P A , Coutinho A
Published: December 2022

Anti-idiotypic antisera (a-Id) were prepared in SJL mice against the dextran B512 (Dex)-specific hybridoma antibody D.16.6 (mu, kappa) of C57BL/6 origin. Such a-Id were specific for the D.16.6 antibody, but the idiotypic determinants recognized were not hapten modifiable. The a-Id were used in radioimmunoassays (RIA) to study the genetic control of the expression of the D.16.6 idiotype (Id) in a variety of mouse strains. It was found that all mouse strains tested expressed this Id as a component of the normal background immunoglobulin (Ig) in their sera. Only in mice of the IgCHb haplotype, however, was the Id found in association with Dex-specific antibodies. Furthermore, even in IgCHb mouse strains, not all individuals expressed Id-positive anti-Dex antibodies. Because the D.16.6 Id is a "recurrent" natural Id, but a "non-dominant" Id in the Dex response of IgCHb mouse strains, these observations contradict our previous suggestions on the selection of "dominant" Id from the pool of "naturally" produced Ig.
mIgM:mIgD ratios on B cells: mean mIgD expression exceeds mIgM by 10-fold on most splenic B cells.
Authors: Havran W L , DiGiusto D L , Cambier J C
Published: December 2022

The relative frequency of mIgM and mIgD molecules on B cell surfaces is important in determining, in large part, the isotype involvement in antigen binding and signal transduction. Although it is generally assumed that on most mature B cells, mIgM and mIgD occur in roughly equal quantities, no formal analysis of this question has been reported. In this report, we describe such an analysis based on the quantitation of anti-Fab or anti-kappa specific immunofluorescence of splenic B cells before or after capping with rabbit anti-IgD or anti-IgM antibodies or both. The results indicate that, whereas mean expression of IgD exceeds IgM on splenocytes by threefold, members of the major B cell subpopulation (60 to 70% of cells) express 10-fold more IgD than IgM.
A genetically restricted suppressor factor that requires interaction with two distinct targets.
Authors: Aoki I , Usui M , Minami M , Dorf M E
Published: December 2022

We have previously described a genetically restricted suppressor factor (TsF3) that suppresses the terminal phases of the contact sensitivity response. The activity of TsF3 is restricted by genes in the H-2 (I-J) and Igh complexes. This report analyzes the mechanisms responsible for these genetic restrictions. One cellular target of TsF3 is an I-J-bearing antigen-presenting cell population that is sensitive to low doses of cyclophosphamide. To elicit suppression I-J homology is required between this antigen-presenting cell population and the TsF3 donor. In contrast, the Igh-linked genetic restriction exists between TsF3 and an unprimed cell population present in the recipient. These findings suggest that under these experimental conditions TsF3 acts by bridging the APC with cells of the host. Finally, we demonstrated that nonspecific bystander or cognate suppression can be mediated by TsF3, provided specific antigen is present in the site of the ongoing T cell response.
Circulating lymphocyte subpopulations in pregnancy: a longitudinal study.
Authors: Tallon D F , Corcoran D J , O'Dwyer E M , Greally J F
Published: December 2022

Peripheral blood lymphocyte subpopulations were studied sequentially during the gestation and the postpartum period of 84 obstetrically normal women. Both the percentage and absolute numbers of total peripheral blood lymphocytes decreased in the second and third trimester. Although there was a further decrease in percentage numbers 1 wk postpartum, the absolute numbers returned to normal. Absolute numbers of OKT3+ and of OKT4+ cells decreased in the second and third trimesters; OKT8+ cells decreased in the third trimester only. Absolute numbers of all T cells returned to normal postpartum. The absolute numbers of B lymphocytes, while showing no consistent pattern, decreased significantly in the third trimester and in the second postpartum period; however, the percentage values of all lymphocyte subpopulations did not alter during or after pregnancy. The data do not support the concept that a numerical imbalance in the T cell subpopulations has a causative role in the immunodeficiency of pregnancy.
Immunological studies of aging. IX. Impaired proliferation of T lymphocytes detected in elderly humans by flow cytometry.
Authors: Staiano-Coico L , Darzynkiewicz Z , Melamed M R , Weksler M E
Published: December 2022

Lymphocytes from humans over the age of 65 incorporate approximately 50% less tritiated thymidine than do lymphocytes from young donors when cultured with phytohemagglutinin. Because lymphocytes from elderly humans are more sensitive to cell cycle arrest induced by tritiated thymidine, it was impossible to determine to what extent impaired thymidine incorporation reflected a defect in proliferation or the increased sensitivity to the radioactive isotope. Flow cytometry was used to measure the proliferative response of T cells from young and old donors in culture with PHA. It was found that 25 percent fewer lymphocytes from old as compared to young humans enter the G1 or complete the S phase of the cell cycle. However, the rate of progression through the cell cycle by activated cells from young and old humans is comparable. Thus, flow cytometry suggested that the difference in thymidine incorporation by lymphocytes from old and young donors is attributable equally to a proliferative defect and to cell cycle arrest induced by tritiated thymidine. This conclusion was supported by the fact that the relative impairment of thymidine incorporation by lymphocytes from old donors was only one-half as great when a 20-min instead of a 24-hr pulse of tritiated thymidine was used.
Functional status of cells from lymphoid and myeloid tissues in mice with severe combined immunodeficiency disease.
Authors: Dorshkind K , Keller G M , Phillips R A , Miller R G , Bosma G C , O'Toole M , Bosma M J
Published: December 2022

Cells from mice with severe combined immunodeficiency disease (SCID) were tested in assays that measure myeloid and lymphoid function. Results showed that C.B-17 scid and their normal counterparts (C.B-17) have similar levels of spleen colony-forming units. The frequency of in vitro myeloid colony-forming units in C.B-17 scid spleen is elevated, but the absolute number of colony-forming units in C.B-17 scid and C.B-17 spleen is similar. The absolute number of bone marrow colony-forming units in C.B-17 scid and C.B-17 mice is comparable. Cells from C.B-17 scid spleen are consistently negative in all tests of B and T cell function. C.B-17 scid splenocytes fail to proliferate in response to T and B cell mitogens or to allogeneic lymphocytes in a one-way MLR; C.B-17 scid cells do serve as stimulators in MLR. B lymphocyte colony-forming units are absent, as are cytotoxic lymphocyte precursors and cells that can generate T cell colonies with cytotoxic progenitors. The microenvironment of the C.B-17 scid mouse is conducive to lymphocyte differentiation, because functional B and T cells are easily detectable in mice reconstituted with normal bone marrow cells. The results of this study indicate that scid specifically impairs the differentiation of stem cells into mature lymphocytes; myeloid cell differentiation is not affected.
Genetic analysis of susceptibility to Theiler's virus-induced demyelinating disease in mice.
Authors: Lipton H L , Melvold R
Published: December 2022

Genetic control of resistance and susceptibility to Theiler's murine encephalomyelitis virus (TMEV)-induced demyelinating disease in mice was examined. Mice derived from various crosses between susceptible SJL (S) and resistant C57BL/6 (B6) strains allowed determination of the relative susceptibility contributed by H-2 and non-H-2 genes, as well as allowing segregation of H-2 linked genes. Six of 39 B6SF1 hybrid progeny developed disease, suggesting several possibilities--resistance is dominant and susceptibility recessive but with incomplete penetrance, the incidence of disease is dose dependent and is dictated by the number of "susceptible/resistant" alleles present at one or more loci, or some combination of the two. Data involving the progeny from the B6SF1 hybrids mated to the S parental strain indicated the involvement of more than one locus. The strong correlation between the number of H-2s haplotypes and the incidence of demyelinating disease suggested that at least one of the genes was linked to the H-2 complex; however, other independently segregating (unlinked) loci strongly affected the disease incidence among H-2 identical animals. A gene or genes from the S background was also associated with enhanced virus growth or diminished virus clearance in the central nervous system (CNS). Overall, the observations support a gene dosage model for susceptibility that relates disease incidence to the total number of S alleles at both H-2 and non-H-2 loci.
Acetylated diamines inhibit endotoxin-induced lymphocyte activation.
Authors: Ryan J L , Bondy P K , Gobran L , Canellakis Z N
Published: December 2022

Fully acetylated diamines hexamethylenebisacetamide (HMBA) and diacetylputrescine (tetramethylenebisacetamide, TMBA) cause inhibition of murine B lymphocyte activation in the absence of ruminant serum. Mitogenesis induced by LPS, LAP, and 8-BrcGMP was profoundly inhibited in the presence of 3 mM HMBA and TMBA. PHA-stimulated thymidine uptake was not affected. Higher levels of the acetylated polyamines were toxic to lymphocytes. Immunoglobulin secretion by LPS-stimulated spleen cells was inhibited in similar fashion. This inhibition requires the early addition of the polyamines and does not appear to be associated with direct inhibition of DNA synthesis. These data support the hypothesis that polyamines represent immunomodulatory agents and suggest that acetylated derivatives of polyamines may also have activity in regulating B lymphocyte activation.
Functional studies on the secreted form of human C4 (C4s), two incompletely processed two-subunit C4 molecules (beta - alpha + gamma and beta + alpha - gamma), and pro-C4.
Authors: Chan A C , Atkinson J P
Published: December 2022

The functional properties of the secreted form of C4 (C4s), which has a Mr approximately 5000 greater than the predominant C4 molecule found in plasma (C4p), two incompletely processed two-chain C4 molecules (beta - alpha + gamma and beta + alpha - gamma), and the extracellular C4 precursor (designated pro-C4(E)] were evaluated. All four molecules are secreted in parallel by a human hepatoma-derived cell line (Hep G2). Secretion of hemolytically active C4 is linear up to approximately 12 hr, peaks at 24 hr, and then progressively decreases over the next 48 hr. This loss of C4s functional activity parallels the proteolytic conversion of C4s to C4bs. To compare the hemolytic efficiencies of C4s and C4p, a solid-phase competitive radioimmunoassay was developed to permit measurement of the small quantities of C4 antigen in these cultures. The hemolytic efficiencies of C4s and C4p were similar. These results indicate that extracellular processing of C4s to C4p does not modulate the hemolytic activity of the molecule. Consistent with their ability to bind methylamine, both the alpha s-chain and the alpha - gamma subunit undergo denaturation-induced autolysis. The extracellular and intracellular pro-C4 molecules are also sensitive to autolytic cleavage. Interestingly, the beta - alpha subunit is resistant to autolysis. In experiments in which C4s and C4p were cleaved by C1-s to C4bs, C4(beta - alpha + gamma), C4(beta + alpha - gamma), and pro-C4(E) were resistant to C1-s cleavage and thus hemolytically inactive relative to C4s. These data indicate that processing of C4 to a three-chain structure is required to provide the proper conformation for efficient activation by C1.
IgE immune complexes induce immediate and prolonged release of leukotriene C4 (LTC4) from rat alveolar macrophages.
Authors: Rankin J A , Hitchcock M , Merrill W W , Huang S S , Brashler J R , Bach M K , Askenase P W
Published: December 2022

Alveolar macrophages obtained by lung lavage from rats were incubated with monoclonal mouse anti-DNP IgE and specific antigen (DNP-HSA) and were found to release a slow reacting substance (SRS), which was characterized by high performance liquid chromatography as leukotriene C4 (LTC)4. Alveolar macrophages incubated with 1 microM A23187 (calcium ionophore) released similar amounts of SRS (6.0 +/- 2.2 and 5.7 +/- 3.7 X 10(-10) mol of LTC4 per 5 X 10(6) alveolar macrophages, respectively). The optimal conditions and mechanism of LTC release by IgE and antigen were examined. LTC4 release was maximal when freshly retrieved alveolar macrophages were incubated for 20 min with 10 micrograms/ml IgE and then for 20 min with 100 ng/ml antigen or for 20 min with IgE and antigen that had been preincubated together for 30 min at room temperature. In addition, LTC4 release was maximal when cells were challenged with IgE and antigen in a protein-free balanced salt solution and when the cells were tumbled to prevent adherence. Dose response experiments revealed that macrophages released LTC4 when stimulated with as little as 10 ng IgE and 100 ng DNP-HSA. Alveolar macrophages did not release LTC when challenged with IgE or DNP-HSA alone. Activation of LTC4 release by IgE and antigen was rapid in onset (2.5 to 5 min), and washing to remove fluid phase IgE and antigen revealed that once activated, alveolar macrophages were capable of prolonged and continuous release of LTC4. Peritoneal lavage cells stimulated with IgE and antigen did not release SRS but could release SRS when incubated with A23187 (5.7 +/- 1.3 X 10(-10) mol LTC4/5 X 10(6) macrophages). A large variability existed between individual rats in the ability of their alveolar macrophages to be activated by IgE and antigen to release LTC4. DNP-HSA labeled with 125I was used to show formation of immune complexes of IgE and antigen when IgE and antigen were incubated together before macrophage challenge. IgE immune complexes containing as little as 2 ng of antigen elicited the release of LTC4 from alveolar macrophages. These data indicate that rat alveolar macrophages release primarily LTC4 when challenged with IgE immune complexes, and that the alveolar macrophage may differ in this respect from peritoneal macrophages that do not release detectable quantities of LTC4 when challenged under identical conditions.
Suppression of human lymphocyte responses by oral spirochetes: a monocyte-dependent phenomenon.
Authors: Shenker B J , Listgarten M A , Taichman N S
Published: December 2022

Spirochetes have been implicated in the pathogenesis of several human infections including syphilis, yaws, Lyme disease, and periodontal diseases. We examined soluble sonic extracts of oral spirochetes (Treponema denticola and T. vincentii) for their ability to alter human lymphocyte function. These organisms were isolated from subgingival plaque of patients with periodontitis. We found that sonicates of several but not all strains of T. denticola caused a dose-dependent inhibition of human lymphocyte responsiveness to Con A, PHA, PWM, and the recall antigen SKSD. Suppression involved alterations in DNA, RNA, and protein synthesis; there was no effect on cell viability. In contrast, sonicates of T. vincentii (medium-sized spirochetes) had no demonstrable effects on lymphocyte activation. The suppressive factor derived from T. denticola is heat-labile with a m.w. of approximately 100,000. To achieve maximal suppression, sonicates had to be added during the first 24 hr of incubation; there was no inhibition observed when added at 48 or 72 hr (along with 3H-TdR). Suppression was dependent on the presence of adherent monocytes; removal of these cells prevented spirochete-induced suppression of lymphocyte proliferation. Furthermore, the combination of indomethacin and catalase were able to reverse (or prevent) the inhibitory effects of the spirochete extracts, demonstrating a requirement for both prostaglandins and hydrogen peroxide. The potential role of such suppressive factors in periodontal disease is discussed.
Cutaneous leishmaniasis in anti-IgM-treated mice: enhanced resistance due to functional depletion of a B cell-dependent T cell involved in the suppressor pathway.
Authors: Sacks D L , Scott P A , Asofsky R , Sher F A
Published: December 2022

The contribution of B cells and antibodies to either the resistance or susceptibility to cutaneous leishmaniasis has been investigated in mouse strains rendered B cell-deficient by treatment with anti-mouse IgM antisera from birth (mu-suppressed). These studies confirm that immunity to cutaneous disease in a normally resistant mouse strain (C3H/HeJ) is independent of antibody, but that B cells and/or antibodies are required for the evolution of suppressed DTH and the consequent disease susceptibility of BALB/c mice. Anti-IgM-treated BALB/c mice, which lacked detectable anti-leishmanial antibodies during the course of infection, displayed a sustained DTH response to leishmanial antigen and were able to control their cutaneous lesions. The enhanced resistance of mu-suppressed mice could be completely abrogated by transfer of suppressor T cells from infected control animals into mu-suppressed mice before their infection. Thus the suppressor T cells, which are generated during leishmanial infection in BALB/c mice, can effect suppression in the absence of antibody. Evidence that B cells or antibodies are required for the generation of suppressor T cells was demonstrated by using BALB/c mice in which suppressor T cells fail to be generated during infection as a result of prior sublethal irradiation. Splenic T cells from normal mice could overcome the resistance conferred by sublethal irradiation, whereas splenic T cells from mu-suppressed mice could not. Thus the enhanced resistance of mu-suppressed BALB/c mice appears to be a consequence of their lack of functional expression of a B cell-dependent T cell critical to the suppressor pathway.
Genetic control of resistance to Listeria monocytogenes: regulation of leukocyte inflammatory responses by the Hc locus.
Authors: Gervais F , Stevenson M , Skamene E
Published: December 2022

The control mechanisms responsible for the innate resistance of C57BL/6J (B) mice and for the innate susceptibility of A/J (A) mice to infection with Listeria monocytogenes were studied by typing the recombinant inbred (RI) strains derived from these two progenitors for the trait of Listeria resistance/susceptibility. The strain distribution pattern (SDP) of this trait obtained in the 13 AXB/BXA RI strains studied suggests that an allelic difference at a major locus (Lr-1) is responsible for the trait of resistance/susceptibility to Listeria. In addition, another putative gene (Lr-2) unlinked to Lr-1 is postulated to control the level of bacterial load within the group of susceptible strains. The SDP of A and B alleles at the Lr-1 locus was fully concordant with that observed for the Hc locus (controlling the level of the C5 component of complement). This suggests that the genetic susceptibility of A strains of mice to Listeria infection is either directly due to or related to the C5 deficiency found in that strain. This conclusion is enhanced by the observation of a significant protection of A mice from listeriosis by the infusion of C5-rich serum. A survey of RI strains for the magnitude of PMN and macrophage inflammatory responses showed that the expression of both traits co-segregated and that the C5 deficiency was the major factor responsible for the defective inflammatory response of A strain mice. We conclude that a defect in the phagocyte inflammatory responses caused by C5 deficiency is the major reason for the extreme susceptibility of A mice to Listeria.
Tumor radioimmunolocation: differential antibody retention by antigenic normal tissue and tumor.
Authors: Ballou B , Jaffe R , Taylor R J , Solter D , Hakala T R
Published: December 2022

On the assumption that a high specificity would be required for in vivo tumor location and drug targeting, much labor has been expended to produce monoclonal antibodies directed to antigens found primarily on tumors. However, for at least one tumor-associated antigen, stage-specific embryonic antigen 1 (SSEA-1), the concentration of the target antigen need not be higher in the tumor than that found in normal tissues for successful use of the corresponding monoclonal antibody in in vivo tumor location; rather, accessibility and antibody metabolism are more important.
The systemic administration of purified interleukin 2 enhances the ability of sensitized murine lymphocytes to cure a disseminated syngeneic lymphoma.
Authors: Donohue J H , Rosenstein M , Chang A E , Lotze M T , Robb R J , Rosenberg S A
Published: December 2022

Significant serum titers of interleukin 2 (IL 2) can be maintained in mice for 12 h after i.p. injection in a 15% gelatin solution. We have tested the ability of IL 2 administered systemically in this fashion to enhance the therapeutic effect of adoptively transferred specifically sensitized lymphoid cells that were expanded in IL 2. Mice with established local and disseminated FBL-3 lymphoma, induced by intrafootpad injection of 10(7) cells after 500 rad total body irradiation, were treated with a combination of i.v. injected murine splenocytes and either murine supernatants containing IL 2 or pure human IL 2 in gelatin. Splenocytes from immune mice were resensitized to irradiated tumor in vitro and were expanded for 7 days in lectin-free IL 2 supernatants. Treatment with these murine splenocytes administered with murine IL 2 supernatants prolonged mean survival to 33.6 days compared with mean survival times of 16.9 days (p less than 0.001) and 23.4 days (p = 0.007) for mice treated with IL 2 alone or splenocytes alone. Human IL 2, purified to homogeneity from the Jurkat cell line, was also capable of improving the therapeutic efficacy of transferred cells in mice. Mean survival was significantly prolonged to 32.1 days when cells and purified human IL 2 were administered, whereas mean survival times of 18.1 days (p = less than 0.001) and 21.5 (p = less than 0.001) were seen for mice treated with IL 2 alone or expanded immune cells alone. Cure rates in this model were also significantly enhanced with the combined treatment of IL 2 and expanded immune cells. Combined immunotherapy utilizing IL 2 and immune cells was thus significantly better than either component used alone. The systemic administration of IL 2 in conjunction with sensitized expanded lymphoid cells may be a useful approach to the immunotherapy of other murine and human tumors.
LFA-1, LFA-2, and LFA-3 antigens are involved in CTL-target conjugation.
Authors: Krensky A M , Robbins E , Springer T A , Burakoff S J
Published: December 2022

Three cell surface antigens associated with the CTL-target cell interaction were previously identified by generation of mAb against OKT4+, HLA-DR-specific CTL, and selection for inhibition of cytolysis in a 51Cr-release assay. In this report, we showed that these mAb inhibit cytolysis by blocking CTL-target cell conjugate formation. It appears that LFA-1, LFA-2, and LFA-3 are cell surface structures involved in strengthening effector-target adhesion that accompanies antigen-specific recognition.
Generation of the alloreactive T cell repertoire: K region homology between H-2b T cell precursors and T cell maturation environment is required for the generation of the Kbm6-specific cytotoxic T cell repertoire.
Authors: Gress R E , Hodes R J
Published: December 2022

The influence of T cell genotype and T cell maturation environment on the generation of the T cell alloreactive repertoire was evaluated in the H-2b cytotoxic T lymphocyte response to Kb mutant determinants expressed by the strain B6-H-2bm6. Specifically, by constructing radiation bone marrow chimeras with B6 or B10 (H-2b) donor cells and B10.BR, B10.A(4R), B10.MBR, and B6.C-H-2bm1 irradiated mice as recipients, it was possible to investigate the major histocompatibility complex (MHC)-encoded gene products of the host environment required for the generation of a bm6-specific H-2b CTL response. The results of such experiments confirmed the previous finding that the alloreactive T cell repertoire is influenced both by T cell MHC genotype and by the MHC gene products of the T cell maturation environment. In addition, the results of the present study further demonstrated that in the chimeric donor and host genetic combinations used, it was both necessary and sufficient that there be a homology of K region-encoded determinants for the generation of a bm6-specific CTL response. Experiments utilizing a mixed responder population of unresponsive B6----B10.D2 spleen cells and responsive Lyt-2 congenic B6.Lyt-2.1 spleen cell suggested that the cellular defect(s) underlying the unresponsiveness of the chimeric cells to bm6-encoded determinants was at the level of the CTL precursor. Together, these findings indicate that an interaction of the K region-encoded gene products of the T cell and its maturation environment play a critical role in the generation of the CTL repertoire specific for bm6 mutant determinants. We discuss here the possibility that this interaction may reflect a requirement that T cells recognize such mutant allodeterminants in association with self restriction elements present on the same mutant K region-encoded molecule.
Qa-like genes defined by CTL analysis of B10.W lines.
Authors: Vucak I , Juretić A , Vidović D , Nagy Z A , Klein J
Published: December 2022

Twelve responder-stimulator combinations of mouse B10.W strains identical at K, D, and class II loci were tested for the generation of cytolytic T lymphocytes (CTL). No primary CTL could be obtained in any of the combinations but in nine combinations CTL were generated after priming in vivo. Six of these CTL are described. They define five antigenic determinants expressed exclusively in a small group of B10.W lines. The determinants appear to be part of the same system that resembles the Qa system originally defined in classic inbred strains. This resemblance rests on the observation that in vivo priming is necessary for the generation of the CTL, and that the CTL are not restricted in their reactivity by known H-2 loci. At least some of the determinants, however, appear to be controlled by a locus (or loci) associated with the K-rather than the D-end of the H-2 complex. Furthermore, some of the CTL directed against these Qa-like determinants cross-react with a molecule controlled by the K locus.
Mechanism of systemic immune suppression by UV irradiation in vivo. II. The UV effects on number and morphology of epidermal Langerhans cells and the UV-induced suppression of contact hypersensitivity have different wavelength dependencies.
Authors: Noonan F P , Bucana C , Sauder D N , De Fabo E C
Published: December 2022

We previously reported that broad band UV radiation or narrow bands of UV (Hbw 3 nm) of wavelengths 250 to 320 nm cause a systemic suppression of contact hypersensitivity (CHS) in mice, observed when the contact sensitizer is applied to a nonirradiated site. To determine if this effect is associated with UV-induced alterations in epidermal Langerhans cell (LC) numbers and morphology, we performed the following study. LC were identified by ATPase staining of EDTA-separated epidermal sheets. Electron microscope studies confirmed that this method was a satisfactory indicator of the presence of LC; we found no evidence for LC which did not stain for ATPase in either irradiated or unirradiated epidermis. Mice were irradiated on the back with narrow band UV of peak wavelength 270, 290, or 320 nm. The irradiated skin was excised 24 hr later and was stained as described. The number of LC with ATPase staining dendrites and the number of nondendritic LC were enumerated. We found that UV radiation of 270 or 290 nm caused 1) an alteration in LC morphology (loss of dendrites) and 2) a decrease in the total number of epidermal LC. Both effects occurred in a dose-dependent fashion. Previously, these same wavelengths of narrow band UV, but at higher doses, had been shown to cause systemic suppression of CHS. In this study, the doses of 270 or 290 nm UV that resulted in the decreased LC numbers and alterations in LC morphology described above were insufficient to cause systemic suppression of CHS. The converse was found if the irradiating waveband of UV had a peak at 320 nm. A dose of 320 nm UV that caused 50% systemic suppression of CHS had no effect on either the number or the morphology of LC at the site of irradiation. In addition, the number and morphology of LC were unaffected in the ventral epidermis (site of contact sensitization) of mice that had been previously irradiated on the back with a systemically suppressive dose of UV. We conclude: (a) UV-induced alterations in the number and morphology of LC at the site of irradiation are not necessary for the generation of systemic suppression of CHS by UV radiation; this indicates that the initial UV-absorbing event triggering systemic suppression is neither a loss of, nor morphologic alterations to, LC at the irradiation site. (b) A systemic effect of UV radiation on the number and morphology of LC at the unirradiated site of contact sensitization does not occur, and thus is not responsible for the UV-induced systemic suppression of CHS by UV radiation.
Recipient contributions to serial passive transfer of experimental allergic encephalomyelitis.
Authors: Wegmann K W , Hinrichs D J
Published: December 2022

EAE can be adoptively transferred into normal recipients by the transfer of BP-specific EAE effector cells. After cell transfer, a series of ill-defined events occur in the recipient that culminate in the development of paralysis associated with neural tissue damage. We investigated the subsequent recipient response to the adoptively transferred disease and examined the role that recipient lymphocytes play in the development of adoptively transferred EAE. Recipient involvement was assessed by the transfer of EAE through a series of normal F1 animals as recipients and at the endpoint of the experiment, determining the MHC restriction pattern of the BP-sensitive cells present. The serial transfer of EAE from BP-CFA-sensitized LEW----(LEW X F-344)F1----(LEW X P2)F1, and from BP-CFA sensitized LEW----(LEW X F344)F1----(LEW X F-344)F1, resulted in the development of BP-sensitive cells in the spleens of the secondary recipients that were able to transfer disease into normal LEW recipients. To test directly for the development of host-derived BP-sensitive cells that might arise in the F1 animal, the serial transfer of EAE from LEW----(LEW X ACI)F1----(LEW X ACI)F1 was performed. When BP-sensitive cells obtained from the secondary (LEW X ACI)F1 recipient animal were transferred into either normal LEW and ACI, or irradiated LEW and ACI animals as final recipients, transfer of disease was successful only into the LEW parental. These results suggest that the development of passive EAE is due solely to the transferred BP-sensitive cells originating from the actively immunized donor, and that no host-derived lymphocytes are recruited into the pool of EAE effector precursor cells found in the spleen of animals after the development of adoptively transferred EAE.
A rat monoclonal antibody that binds specifically to mouse T lymphoblasts and inhibits IL 2 receptor functions: a putative anti-IL 2 receptor antibody.
Authors: Osawa H , Diamantstein T
Published: December 2022

To obtain an anti-interleukin 2 (IL 2) receptor antibody, we immunized rats with phorbol myristate acetate-pulsed mouse T lymphoblasts. The spleen cells of the rats were fused with rat myeloma cells. Several stable clones of hybridoma cells were obtained that produced monoclonal antibodies (mAb) reacting specifically with mouse T lymphoblasts. One of these mAb, AMT-13, showed characteristics of a putative anti-IL 2 receptor antibody. As tested by FACS analysis, the mAb AMT-13 binds to murine T lymphoblasts or to IL 2-dependent T cell lines, but not to B lymphoblasts, thymocytes, splenocytes, lymph node cells, or bone marrow cells. The mAb AMT-13 inhibited in a species-specific and dose-dependent manner: i) the capacity of mouse T lymphoblasts to absorb IL 2 activity, and ii) the capacity of mouse T lymphoblasts to proliferate in response to IL 2. The function of T lymphoblasts of rat origin was not affected by the mAb AMT-13. The time course of the acquisition by mitogen-stimulated spleen cells of the capacity to absorb IL 2 activity was paralleled by that of their capacity to bind the mAb AMT-13. Preliminary biochemical analysis of the antigen recognized by AMT-13 revealed a major component with an apparent m.w. of 50,000 to 60,000. According to these data, the mAb AMT-13 seems to be directed against an antigenic determinant of the murine IL 2 receptor molecule.
Expression of antigens of the 'T200' family of glycoproteins on hemopoietic stem cells: evidence that thymocyte cell lineage antigens are represented on 'T200'.
Authors: Ralph S J , Berridge M V
Published: December 2022

A panel of monoclonal antibodies has been used to investigate the presence of antigens of the 'T200' family of cell surface glycoproteins on hemopoietic stem cells. As measured by spleen colony inhibition using CBA bone marrow, stem cells were shown to be antigenically heterogeneous with respect to 'T200' family antigens; greater than 60% loss of spleen colonies was observed with 74/8 and 30F11 monoclonal antibodies and with Ly5.1 alloantiserum, whereas little effect was observed with 30G12. Two of four monoclonal antibodies with specificity for B220 showed significant inhibition of spleen colony formation. Monoclonal antibodies against Mac-1, another family of cell surface glycoproteins, had little effect on stem cells. In general, a similar pattern of spleen colony inhibition was observed in CBA, BALB/c, and C57/BL/6 mice. Spleen colony inhibition did not correlate with antibody isotype or with antibody binding to thymocytes, B lymphocytes, macrophages, or platelets as determined by indirect radioimmunoassay. Cell lineage antigens held in common between thymocytes and stem cells were shown to be wholly represented on the T200 glycoprotein. Thus, the anti-stem cell activity in anti-thymocyte serum (ATS) was absorbed by BW5147 T200+ cells but not by a T200- variant. The results show that some antigens of the 'T200' family of cell surface glycoproteins defined with monoclonal antibodies are expressed on stem cells and that other T200 antigens represented in ATS but not as yet identified with monoclonal antibodies show T lineage restriction.
Stabilization of the amplification convertase of complement by monoclonal antibodies directed against human factor B.
Authors: Daha M R , Deelder A M , Van Es L A
Published: December 2022

Three IgM mouse monoclonal antibodies (MoAb), 5B5-A, 2D2-B, and 5B12-A, were prepared by fusion of spleen cells from mice immunized with human B with the SP 2/0 myeloma cell line. They were assessed for their effect on cell-bound and fluid-phase amplification convertases of complement (C) with purified proteins in vitro. 5B5-A and 2D2-B were similar in their effects on cell-bound preformed C3bBb in that they bound to cell-bound C3bBb, stabilized the C3bBb convertase, and rendered the C3bBb convertase relatively resistant to the plasma protein H. These two MoAb were also able to enhance C3 consumption in vitro in reaction mixtures containing C3b, C3, B, and D. At the same time, they presumably stabilized the C3 convertase and caused relative sparing of B hemolytic activity in the reaction mixtures. In contrast, 5B12-A, which also bound to Bb and C3bBb, did not induce any stabilization, but rather caused accelerated decay of cell-bound C3bBb. These results indicate that MoAb against B can have C3NeF-like activity. On the other hand, not all MoAb against B have stabilizing activity on the C3bBb convertase.
Cathepsin G in human mononuclear phagocytes: comparisons between monocytes and U937 monocyte-like cells.
Authors: Senior R M , Campbell E J
Published: December 2022

Previous studies demonstrating that a continuous line of human monocyte-like cells (U937) and human monocytes contain elastase probably identical to human neutrophil elastase suggested the possibility that mononuclear phagocytes share other proteinases with neutrophils. The present work establishes that U937 cells contain cathepsin G, an enzyme heretofore found only in neutrophils. U937 cells contain approximately 10 micrograms of cathepsin G-like activity per 10(7) cells, about 25% of the cathepsin G activity in human neutrophils. Normal monocytes have minimal cathepsin G-like activity (approximately 0.1 microgram per 10(7) cells). The cathepsin G-like activity of U937 cells appears to be due to an enzyme that is the same as cathepsin G by several criteria: 1) it is a serine proteinase with activity like cathepsin G against a synthetic chymotrypsin substrate, succinyl-ala-ala-pro-phe-p-nitroanilide; 2) the proteolytic fragments it releases from fibronectin match those released by cathepsin G; 3) like cathepsin G, it can be purified by sequential Trasylol-Sepharose affinity chromatography and carboxymethyl-Sephadex ion exchange chromatography; 4) its amino acid composition and migration on SDS-polyacrylamide gel electrophoresis are indistinguishable from cathepsin G; and 5) it binds with antiserum raised to cathepsin G. The presence of cathepsin G in U937 cells, in much higher concentration than in normal monocytes, indicates either that the content of cathepsin G in monocytes decreases markedly during monocyte differentiation or that U937 cells differ from normal immature monocytes with respect to synthesis of this neutral proteinase.
Phagocytosis inhibits the expression of tissue transglutaminase in mouse peritoneal macrophages.
Authors: Mehta K , Murtaugh M P , Juliano R L , Davies P J
Published: December 2022

Culture of mouse peritoneal macrophages in serum-containing medium produces a rapid and marked induction of tissue transglutaminase. Concurrent endocytosis of a variety of particles via specific and nonspecific routes suppresses or completely abolishes the induction of the enzyme. The suppression is dose-dependent and is reversible. The rate of recovery from the inhibitory effect, however, depends on the nature of the particles internalized. The inhibitory effect is due to the inhibition of the synthesis of tissue transglutaminase.
Monocyte suppression of antigen-specific lymphocyte responses in diffuse cutaneous leishmaniasis patients from the Dominican Republic.
Authors: Petersen E A , Neva F A , Barral A , Correa-Coronas R , Bogaert-Diaz H , Martinez D , Ward F E
Published: December 2022

Patients from the Dominican Republic with diffuse cutaneous leishmaniasis showed in vivo and in vitro anergy to leishmanial antigen. Relatives of these DCL patients living in the same endemic area frequently showed skin test and lymphocyte reactivity to leishmanial antigens. This further supports the concept of specific anergy in patients with diffuse cutaneous leishmaniasis. Adherent suppressor cells modulate the antigen-specific lymphocyte proliferative response. Suppressor cells could also be isolated by Percoll gradient centrifugation. Co-culturing of lymphocytes and monocytes from HLA-identical leishmanin responders and nonresponders also identified the suppressor cell as a monocyte. In one patient, this suppression disappeared when clinical cure had been accomplished.