bluuebunny/crossref_metadata_2025 huggingface dataset:
Control of Vκ Expression in the Mouse
Authors: Claflin J. Latham , Wei Shaio-Ping L.
Published: December 2022

Somatic cell hybrids were produced between a myeloma cell line of MPC11 and spleen cells from AKR and BALB/c mice immunized with the PC-containing bacteria Streptococcus pneumoniae R36A. Among the PC-specific hybridomas examined, 24 of 26 (92%) of BALB/c and 19 of 30 (57%) of AKR expressed idiotypic determinants and L chains characteristic of a major conserved clonotype in mice, the T15 clonotype. This compares favorably with percent of T15 idiotype-positive antibody and B cells found in the anti-PC repertoire of these two strains. Previous studies have described two genetic variants of the T15 clonotype L chain, Igk-Pcb (BALB/c) and Igk-Pca (AKR), which appear to be controlled by an autosomal locus linked to Ly-3. All of the T15-positive BALB/c hybridomas expressed L chains bearing the Vκ marker, IgK-Pc-B as expected. All of the T15-positive AKR hybridomas, except one, expressed the IgK-Pc-A marker. The exception was a hybridoma, F10-1 expressing a T15 clonotype L chain, Pc-“B,” whose electrophoretic and antigenic characteristics were indistinguishable from Pc-B. The implication of this finding is discussed in relation to known examples of nonallelic behavior of immunoglobulin allotypes.
Relationship between Lymphocyte Proliferation and Tumor-Specific Cytotoxicity after Immune RNA Treatment
Authors: Wang Bosco Shang , Mannick John A.
Published: December 2022

Normal mouse lymphocytes were converted to effector cells cytolytic to syngeneic mouse tumor cells in vitro by incubating these lymphocytes with immune RNA (I-RNA) extracted from the lymphoid tissues of guinea pigs immunized with the same mouse tumor. This effect was tumor specific, RNase sensitive, and DNase and pronase resistant. These I-RNA-incubated lymphocytes were also able to undergo proliferation when they were co-cultured with specific tumor cells in tissue culture. Treatment with mitomycin C completely inhibited the blast transformation of I-RNA-incubated lymphocytes in response to specific tumor antigen, whereas the cytotoxic activity of mitomycin C-treated lymphocytes was not affected. There was no significant alteration in cytotoxic capability when lymphocytes were cultured for 0, 24, 48, or 72 hr after I-RNA incubation before testing. The time necessary for I-RNA-incubated lymphocytes to kill specific target cells was not different whether the lymphocytes had been cultured for 48 hr or tested immediately after I-RNA treatment. These results suggest that a proliferative step is not necessary after I-RNA treatment for lymphocytes to manifest tumor-specific cytotoxicity and that I-RNA may act upon at least two different sets of lymphocytes; one undergoing cellular proliferation in response to specific tumor antigen and the other manifesting tumor-specific cellular cytotoxicity.
Macrophage Restriction of Marek's Disease Virus Replication and Lymphoma Cell Proliferation
Authors: Lee Lucy F.
Published: December 2022

Macrophages are shown to restrict the replication of Marek's disease virus (MDV) and isotope uptake by spleen cells from chickens bearing Marek's disease (MD) tumors. The titer of virus from duck embryo fibroblasts (DEF) co-cultivation with MDV-spleen cells pretreated to deplete marcophages was 4- to 18-fold higher than with untreated cells. Treated MDV-spleen cells increased isotope uptake by 2-fold. These restrictive activities are attributable to macrophage regulation of cell proliferation.
Cellular and Humoral Immune Reactivity to Tumor-Associated Antigens in Chickens Infected with Rous Sarcoma Virus
Authors: Hall Michael R. , Qualtiere Louis F. , Meyers Paul
Published: December 2022

With a leukocyte blastogenesis assay and 3 M KCl extracts of chicken cells infected or infected and transformed by various avian tumor viruses (ATV), the time course of development of cellular reactivity in RSV tumor-bearing chickens to antigens expressed on these cell types was examined. Leukocyte reactivity to mitogens in RSV-injected birds was also investigated. Similarly, humoral responses to ATV-antigens and virus-induced antigens were assessed over time, by measuring the development of antiviral neutralizing antibody, as well as antibody to cell surface antigens on various cell types, by means of an isotopic antiglobulin test (IAT). There were no differences in phytohemmaglutinin responsiveness between the leukocytes of RSV-injected birds and control birds, but responses to pokeweed mitogen were sometimes elevated in RSV-injected animals. Cellular immunity to 3 M KCl extracts, principally to those extracts containing structural components of the virus envelope, developed between 3 and 5 weeks post-injection, during tumor growth and regression, and then declined, although the immune response could be restimulated by virus rechallenge. For the most part, responses to nonvirion tumor-specific surface antigen (TSSA) was weak except after RSV rechallenge. Humoral reactivity as measured by IAT also seemed to correlate well with the presence of virus envelope antigens (VEA) on infected indicator cells and appeared before antiviral neutralizing antibody. The data indicated that, at least during the primary response, major immune reactivity in RSV-injected birds appeared to be directed against VEA and that TSSA appeared to be most immunogenic when VEA was also present.
PPD-Induced Immunoglobulin Production in Human Peripheral Blood Lymphocytes
Authors: Nishikawa Shin-Ichi , Hirata Takeo , Izumi Takateru
Published: December 2022

We investigated the relationship between PPD-reactive helper T cells and PWM-reactive helper T cells in polyclonal Ig production of human PBL. Elimination of PPD-reactive T cells by BUdR + light treatment resulted in a loss of helper function in PPD-induced Ig production, but had no effect on helper function in PWM-induced Ig production. On the other hand, elimination of PWM-reactive T cells resulted in a loss of helper function in both PPD-induced Ig production and PWM-induced Ig production.

A blast cell-enriched fraction that was generated by PPD and separated by the velocity sedimentation method contained helper function in both responses. On the other hand, blast cell-depleted fractions did not contain PPD-reactive helper function, although the PWM-reactive helper function was evident.

These results strongly suggest that PPD-reactive helper T cells are included in PWM-reactive helper T cells.
The Cytolytic T Lymphocyte Response to Trinitrophenyl-Modified Syngeneic Cells
Authors: Finberg Robert , Greene Mark I. , Benacerraf Baruj , Burakoff Steven J.
Published: December 2022

Mice were primed subcutaneously with trinitrophenyl (TNP)-modified syngeneic spleen cells. Seven days later, spleen cells from these in vivo primed mice, or spleen cells from naive mice, were co-cultured with TNP-modified syngeneic cells. Spleen cells from the in vivo primed mice demonstrated augmented cytolytic T lymphocyte (CTL) activity. The spleens of these in vivo primed mice contained a population of radioresistant, antigen-specific, helper T cells. Specifically, spleen cells from these mice, after x-irradiation, were able to augment the in vitro CTL response of normal spleen cells to TNP-modified syngeneic cells.
Restriction of Complement-Mediated Membrane Damage by the Eighth Component of Complement: A Dual Role for C8 in the Complement Attack Sequence
Authors: Nemerow Glen R. , Yamamoto Ken-Ichi , Lint Thomas F.
Published: December 2022

Inhibitors of the fluid phase complex were examined in serum from which the lipoproteins had been removed. A limited number of these nonlipoprotein inhibitors were observed, and one was shown to be C8 by several criteria. The inhibitory activity exhibited numerous physiochemical similarities to C8 including identical chromatographic profiles on DEAE-cellulose, CM-cellulose, and QAE-Sephadex ion exchange columns as well as an indistinguishable m.w. as determined by molecular-sieve chromatography and sucrose gradient ultracentrifugation. In addition, serum from a patient genetically deficient in C8 was shown to lack the inhibitory activity that co-purified with C8 in normal serum.

Subsequent purification of C8 by established methods verified that these activities were indeed due to the same molecule and furthermore that the inhibitory activity was not due to inactive C8, thus establishing a new function for this C component. A comparison of the inhibitory activity of C8 with its hemolytic effeciency revealed that very small amounts were required for both activities, although higher amounts of C8 were required for inhibition of formation than for lysis of preformed EAC1-7 in the presence of excess C9.

Experiments employing purified and radiolabeled C7 indicated that C8 limited the formation of intermediates by preventing the attachment of the fluid phase complexes to target membranes. In addition, the inhibitory activity of C8 could be counteracted by the simultaneous addition of C56̅ and C7 but not by the addition of either component alone, verifying that C8 restricts formation by interacting with the nascent complex in the fluid phase. The ability of C8 to limit the attachment of fluid phase , while being required for the lysis of cells already bearing , suggests a dual role for C8 in complement-mediated cytolysis and thus represents a point of internal regulation in the complement attack sequence.
Immunity to the T1699 Murine Mammary Tumor
Authors: Parthenais Elaine , Haskill Stephen
Published: December 2022

The thymus dependency of humoral immunity to syngeneic tumor antigens was investigated in T cell-deficient DBA/2 mice. ATXBM animals bearing the T1699 mammary tumor in the subcutaneous abdominal area displayed normal immediate but not delayed hypersensitivity reactions (DHR) to 3 M KCl-extracted T1699 antigens injected into the footpad. Sera from ATXBM tumor-bearers passively transferred immediate hypersensitivity but failed to support tumor-specific macrophage-mediated ADCC reactions. The synthesis of macrophage-mediated ADCC antibody was greatly reduced in the CXBM animals when compared to nonirradiated tumor-bearers. The CXBM mice, however, showed normal T cell function as measured by allograft rejection, antibody response to sheep erythrocytes, and DHR to T1699 antigens. Of all antibody classes and subclasses tested by indirect membrane fluorescence, only IgG2b was found to be produced at normal levels by either ATXBM or CXBM tumor-bearers. The results show that IgG2b antibody production in response to T1699 syngeneic tumor antigens is thymus independent and suggest that this antibody is the mediator of immediate hypersensitivity. The synthesis of macrophage-mediated ADCC antibody (IgG2a) was found to be not only thymus dependent but also sensitive to the long-term effects of irradiation and bone marrow repopulation.
Immunochemistry of Salmonella O-Antigens: Specificity of Rabbit Antibodies against the O-Antigen 4 Determinant Elicited by Whole Bacteria and O-Antigen 4 Specific Saccharide-Protein Conjugates
Authors: Jörbeck Hans J. A. , Svenson Stefan B. , Lindberg Alf A.
Published: December 2022

In two precipitating systems using the O-antigenic polysaccharide (PS) from Salmonella typhimurium as antigen and BO antiserum (anti-O4 and -O12 specificities) or O4 antiserum, the octasaccharide

was the best inhibitor. In order of decreasing effectiveness followed:

.

On a molar basis the relative inhibiting activities were 100:45:12.5:0.3:0.1 (BO antiserum) and 100:40:8:0.3:0.1 (O4 antiserum). These data established that the combining sites of antibodies raised by the native bacterial O4 antigen recognize a structure larger than the tetrasacharide but equal to or smaller than the octasaccharide. When the S. typhimurium PS was precipitated with antiserum elicited by immunization with the octasaccharide (see above) conjugated to bovine serum albumin (Os-BSA) the relative inhibitory activities of four of the saccharides (not including methyl-β-abequoside) were 100:30:1.6:0.1. Thus, the combining sites of antibodies elicited by the Os-BSA conjugate recognized structures close or equal to those of antibodies raised by whole bacteria (BO and O4 antisera). When the S. typhimurium PS was precipitated with an antiserum elicited by immunization with 3-O-α-abequopyranosyl-D-mannopyranose covalently linked to BSA (AM-BSA) the relative inhibitory activities of the five saccharides were 7.5:17:100:0.6:0.2. Hence, the AM-BSA antibodies had about fifteen times higher affinity for the methyl 3-O-α-abequopyranosyl-α-D-mannopyranoside than for the octasaccharide. The importance of using different sized haptens, e.g., di- or octasaccharides, to elicit antibodies useful for diagnostic or immunoprophylactic purposes is emphasized.
Suppression of Natural Cytotoxicity by Spleen Cells of Hydrocortisone-Treated Mice
Authors: Hochman Paula S. , Cudkowicz Gustavo
Published: December 2022

The administration of hydrocortisone acetate (Hc) to (C57BL/6 × C3H)F1 or BALB/c mice by two daily i.p. injections (2.5 or 3.75 mg each) caused a substantial but transient reduction of splenic natural killer (NK) cell activity against YAC-1 lymphoma targets. This effect was not dependent on the thymus, since splenic NK activity declined in congenitally athymic BALB/c.nu/nu mice. The cytotoxic activity lowered in vivo by Hc could be partially restored in vitro simply by removing a subpopulation of cells adherent to, or phagocytic for, carbonyl particles. Thus, NK cells were reversibly suppressed in the spleens of Hc-treated mice. Despite removal of suppressor cells by carbonyl iron, filtration of splenocytes through Sephadex G-10 columns was ineffective. Moreover, the plastic-adherent, phagocyte-rich subpopulation of splenocytes was not enriched with suppressors. NK cells from untreated mice were suppressed in vitro by irradiated (2000 rads of γ-rays) and nonirradiated splenocytes of Hc-treated donors, and by cell-free superatants in which such cells were incubated.

It thus appears that Hc activated presuppressor spleen cells to become suppressors within 48 hr. The paradox of suppressor cell removal by carbonyl iron but not by other adherence techniques does not allow characterization of such cells as to macrophage or lymphoid (pre-T or non-T) origin.
Adoptive Immunotherapy of Leukemia in the Rat, without Graft-vs-Host Complications
Authors: Westcott Owen , Dorsch Susan , Roser Bruce
Published: December 2022

PVG rats bearing a transplantable T cell leukemia were treated with large inocula of lymphoid cells from AUG rats sensitized either against the leukemia or against PVG lymphocytes. AUG and PVG bear identical Ag-B antigens but differ at minor loci, including the Pta loci, which code for differentiation antigens expressed only on peripheral T lymphocytes. Treatment with AUG cells immune to either the PVG leukemia or normal PVG cells resulted in prolonged survival of leukemia rats, a profound but ephemeral leukopenia and prolonged disappearance of leukemic cells from lymphoid tissue. All treated animals, however, eventually died with large, discrete deposits of leukemic cells in both hard and soft tissues. Despite the deliberate mismatching of host and donor cells for minor transplanation antigens, no evidence of GVH symptoms was observed in treated rats. This was interpreted as a result of directing the adoptive immune response to antigens of restricted distribution, i.e., on leukocytes and not on somatic cells.
Immune Response to Levan
Authors: Bona Constantin , Mond James J. , Stein Kathryn E. , House Shirley , Lieberman Rose , Paul William E.
Published: December 2022

Normal adult BALB/c mice immunized with bacterial levan (BL) produce antibodies that bind inulin and BL and that express cross-reactive idiotypes (IdX-G, IdX-A, IdX-B) found on inulin-binding myeloma proteins. In addition, such mice make anti-BL antibodies that fail to bind inulin and that do not express the IdX. Young BALB/c mice immunized with BL or with a variety of inulin conjugates fail to produce antibodies that bind inulin or express IdX although such mice immunized with BL do produce antibodies that bind BL. The late appearance of the capacity to produce IdX+ anti-inulin antibodies is most probably due to a delay in development or maturation of precursor cells rather than to some defect ascribable to the nature of the carrier molecule, to the presence of suppressor cells, or to a microenvironmental defect in the neonate. Thus, inulin determinants on a variety of carriers fail to stimulate the response of young mice; spleen cells from young mice transferred to adult C.B20 mice do not respond unless they have been in residence for 7 to 14 days before immunization; T cells from young mice do not suppress the response of adult BALB/c mice and adult spleen cells can mount a response when transferred into 1-week-old recipients. Furthermore, the polyclonal activator Nocardia water-soluble mitogen (NWSM) stimulates the appearance of anti-BL plaque-forming cells (PFC) but not of anti-inulin PFC in spleen cells from mice of 1 to 3 weeks of age, whereas spleen cells from 4-week-old and older mice develop both anti-BL and anti-inulin PFC in response to NWSM. A most provocative finding is the appearance in the serum of our nonimmunized young BALB/c mice of IdX-A+ molecules that lack IdX-G and -B and that do not bind to inulin or BL. The precursors of cells secreting these IdX-A+ molecules are interesting candidates for a “protoclone” from which precursors of IdX-G+, -A+, -B+ anti-inulin antibody-forming cells develop.
Terminal Transferase-Positive Human Bone Marrow Cells Exhibit the Antigenic Phenotype of Common Acute Lymphoblastic Leukemia
Authors: Janossy George , Bollum Fred J. , Bradstock Kenneth F. , McMichael Andrew , Rapson Nicholas , Greaves Melvyn F.
Published: December 2022

Combined immunologic assays for TdT enzyme and membrane markers show that TdT+ cells in nonleukemic human bone marrow carry ALL-associated and Ia-like antigens but no thymocyte markers or surface Ig. These cells could be precursors involved in acute lymphoblastic leukemia of the “common” or non-T, non-B type and in lymphoid blast crisis of Ph' positive chronic myeloid leukemia. A few TdT+, Ia+ cells express cytoplasmic IgM, indicating that some pre-B cells may be TdT positive.
T Suppressor (TG) Lymphocytes Fluctuate in Parallel with Changes in the Clinical Course of Patients with Multiple Sclerosis
Authors: Huddlestone John R. , Oldstone Michael B. A.
Published: December 2022

Aberrant immune responses to several unrelated antigens observed in patients with multiple sclerosis have suggested faulty regulation of immunocompetent cells. Recently human T lymphocytes have been segregated into subpopulations. T lymphocytes that bear Fc receptors for IgG suppress B lymphocyte proliferation and the production of IgG. Here we report studies on the percentage of TG cells in the peripheral blood of 25 patients all of whom met stringent clinical criteria for the diagnosis of multiple sclerosis (MS). Many of these patients were followed for more than 14 months.

In the total group, nine patients experienced definite acute exacerbations of disease followed by periods of remission. The mean percentages ± 1 standard deviation of TG cells found in all MS patients during remission, 16 ± 6 was significantly higher than the mean found in normal subjects, 12 ± 2. In contrast, the mean found in patients experiencing an acute attack, 6 ± 2 was significantly lower than the mean TG level of normal subjects. Moreover, when individual patients were followed through their clinical illness, levels of TG lymphocytes that were low during acute exacerbations invariably increased to relatively high levels with the onset of clinical remission.
Expression of Ia Antigen on Adult and Neonatal B Lymphocytes Responsive to Thymus-Independent Antigens
Authors: Mond James J. , Sehgal Evan , Sachs David H. , Paul William E.
Published: December 2022

Spleen cells from normal adult or neonatal mice were treated with anti-Ia and C and in vitro responses to two thymus-independent (TI) antigens, trinitrophenyl-Brucella abortus (TNP-BA) and TNP-Ficoll were studied. TNP-BA, a TI antigen that is known to stimulate neonatal B cells, was used in these experiments with the intent of stimulating an early appearing population of B lymphocytes, which might be anticipated to express surface immunoglobulin (sIg+) but not Ia. Cells treated with C and antisera with activity to I region determinants coded for by genes in I-A, I-E, or I-E/C were unable to mount significant responses to TNP-BA or TNP-Ficoll. These diminished responses could not be reconstituted with x-irradiated cells as a source of macrophages. When neonatal spleen cells, which allegedly possess a high fraction of Ig+ Ia- cells, were treated with A.TH anti-A.TL, there was a marked impairment in their ability to respond in vitro to TNP-BA. Thus, even primitive TI responses depend largely on cells that have already expressed Ia.

To assess whether spleen cells treated with anti-Ia and C contained small numbers of immunocompetent cells that might not have been detected in in vitro systems employed, we transferred antisera and C-treated (CBA/N × DBA/2)F1 female spleen cells to B cell defective (CBA/N × DBA/2)F1 male mice and challenged the recipients with TNP-Ficoll 7 or 14 days later. Anti-TNP PFC were measured 5 days after antigen challenge. There was no detectable anti-TNP response seen in F1 male mice reconstituted with anti-Ia + C-treated cells whereas cells treated only with C were able to transfer significant responses.

Responses to TNP-Ficoll were shown to be more sensitive to anti-Ia and C treatment than were responses to TNP-BA suggesting that the subset of B cells responding to TI-2 antigens may bear a higher density of surface Ia than does the B cell subset responding to TI-1 antigens.
Murine T Cell Ia Antigens
Authors: Hayes Colleen E. , Bach Fritz H.
Published: December 2022

Recipient (B10.HTT × A.TH)F1 mice were immunized with Con A-stimulated A.TL thymocytes. Antiserum obtained was strongly cytotoxic to A.TL lymph node cells (50 ± 11% lysis) but not to A.TH or B10.HTT lymphocytes. Cytotoxic anti-I-Ak antibody was observed by lysis of B10.A(4R) lymphocytes (47 ± 10%) in the absence of C57BL/10 reactivity. Exhaustive absorption first with B10.A(4R), then with B10.A(5R), left no A.TL reactivity. Antibody to I-Bk determinants was thereby ruled out. Cytotoxic anti-I-Jk antibody was observed by reactivity of B10.A(4R)-absorbed serum on A.TL cells (19 ± 5%), and confirmed by unabsorbed serum lysis of B10.S(9R) (16 ± 5%) in the absence of B10.S lysis and by cytotoxicity on B10.A(5R) lymphocytes (25 ± 6%).

The I-Jk-controlled antigen has been studied in detail. This specificity cannot be attributed to a Qa or Tla-region gene; these gene products have been formally excluded by an appropriate strain choice for antiserum production. Furthermore, Qa or Tla gene products are excluded since DBA/2 lymphocytes cannot absorb cytotoxic reactivity for A.TL, B10.S(9R), or B10.A(5R) cells. This I-Jk antigen is carried by lymph node cells (19 ± 5% lysis) and splenocytes (8 ± 3% lysis), but not by thymocytes (<3%). Separated B10.A(5R) B and T lymphocytes were clearly distinguishable; T cells were lysed (22 ± 3%), B cells were not (<3%). Moreover, absorption of antiserum with increasing numbers of B lymphocytes did not diminish residual cytotoxic activity on T lymphocytes. Expression of the I-J-controlled antigen is confined to the k haplotype and is therefore considered a private specificity. These experiments lead us to conclude that there are T cell I-region-controlled structures, distinct from classical B cell Ia antigens, that may elicit antibody production under appropriate conditions.
Absence of Unexpected H-2 Alloantigens on a Murine Lymphoma
Authors: Fox Robert I. , Weissman Irving L.
Published: December 2022

The chemical nature of the structures recognized on target cell membranes by cytotoxic lymphocytes (CTL) remains unclear. One hypothesis suggests that new H-2 alloantigens may be de-repressed as a consequence of viral infection or transformation. We have attempted to detect the appearance of any “foreign” H-2 antigens on the Moloney virus-induced lymphoma LSTRA (H-2d), by analyzing 125I-labeled LSTRA membranes from detergent-solubilized cells, immunoprecipitated with anti-H-2 or anti-β2 microglobulin sera.

We have previously demonstrated that LSTRA bears H-2Dd and H-2Kd antigens that are indistinguishable from BALB/c antigens in their size, isoelectric properties, and antigenicity. We now extend these studies by showing that: 1) All molecules precipitated by anti-β2 microglobulin sera (other than β2 microglobulin) could be removed by pretreatment with anti-H-2d sera. 2) LSTRA cells were specifically examined for H-2d antigens (reported by others to be on LSTRA cells); no such molecules were detected. 3) H-2 molecules precipitated from 125I-LSTRA membrane with anti-β2 sera were indistinguishable from those obtained from 125I-BALB/c membrane in their appearance on two-dimensional gels, ruling out modifications of existing H-2 molecules or substitutions for them in association with β-2.

We also noted that three different xenoantisera to β-2 microglobulins precipitated selected populations of H-2-β-2 dimers—either H-2Dd + β-2, or H-2Kd + β-2, or both. This phenomenon might be explained by true heterogeneity of β-2 microglobulins, or by selective expression of different determinants on a homogeneous population of β-2 molecules as a result of their differential association with H-2Kd and H-2Dd glycoproteins.

This publication expands our previous work to characterize H-2 antigens on the membrane of LSTRA cells that can serve as targets for H-2 restricted lysis by immune CTL. The data presented here and in our previous work allow exclusion of certain models of the membrane structures that are recognized on the target cell during cell-mediated lysis.
Specific Absorption of T Lymphocytes Committed to Soluble Protein Antigens by Incubation on Antigen-Pulsed Macrophage Monolayers
Authors: Werdelin Ole , Braendstrup Otto , Shevach Ethan M.
Published: December 2022

Monolayers of macrophages (Mø) pulsed with antigen were used as immunosorbents for T lymphocytes from guinea pigs primed to soluble protein antigens. T lymphocytes were cultured on the Mø monolayers for 4 hr, then aspirated and reincubated on a fresh monolayer pulsed with the same antigen for a second and a third step. T lymphocytes so treated were selectively deprived of cells responding in assay for antigen-dependent proliferation against the antigen used for pulsing the absorbing monolayer, but maintained their response to other antigens. The lymphocytes adhering to the Mø of the absorbing monolayer were capable of giving a full response to the antigen used for pulsing the Mø of the monolayers. The proliferative response of F1 T lymphocytes to antigen in association with Mø of either parental strain could be absorbed leaving the response to antigen in association with Mø of the other parental strain. The absorption of the proliferative response was not inhibited by addition of excess soluble antigen to the medium of the absorption culture. Our results indicate that specific guinea pig T lymphocytes responding by proliferation to soluble protein antigens recognize and bind specifically to a complex of Ia antigen and protein antigen at the surface of the Mø.
In Vivo Elimination by Specific Effector Cells of an Established Syngeneic Rat Moloney Virus-Induced Sarcoma
Authors: Fernandez-Cruz Eduardo , Halliburton Barbara , Feldman Joseph D.
Published: December 2022

BN rats immunized subcutaneously with a viral induced tumor (MST) or with a chemical-induced fibrosarcoma (BC5) were donors of immune spleen cells. Samples of immune spleen cells were tested in vitro against MST and BC5 in a 51Cr release assay before culturing and after 7 days of culture with mitomycin C-treated MST and/or BC5 tumor cells (MSTMit, BC5Mit). These spleen cells were infused in vivo i.v. into x-rayed (400 R) and nonirradiated BN recipients that bore a vascularized and progressive (1 to 1.5 cm in diameter) subcutaneous MST or BC5. Spleen cells from untreated BN donor rats were also tested in vitro and in vivo as controls.

Established MST were specifically eliminated by spleen cells immune to MST after culture with MSTMit, but not by spleen cells immune to MST without further culture nor by cultured or uncultured BC5 immune spleen cells and control spleen cells. Also, the growth of BC5 was not affected by MST immune spleen cells cultured for 7 days with MST and/or BC5. Elimination of Moloney sarcoma (MST) in vivo occurred in less than 35 days and was correlated with the generation of cytotoxicity in vitro since only MST immune spleen cells cultured with MSTMit were able to augment significantly their cytotoxic capability in vitro.
Role of Binding Through C3b and IgG in Polymorphonuclear Neutrophil Function: Studies with Trypsin-Generated C3b
Authors: Newman Simon L. , Johnston Richard B.
Published: December 2022

In previous studies of the role of IgG and C3b in opsonization, C3b has not been present on the target particle in the complete absence of other serum proteins. We have developed a system in which C3b alone is present on the particle, using trypsin cleavage of purified C3. Sheep erythrocytes (E) coated with trypsin-generated C3b (ETC3b) efficiently bound to but were not ingested by human neutrophils. Specificity of binding was demonstrated by inhibition of rosette formation with fluid phase C3 or C3b, and by a decrease in rosettes after treatment of ETC3b with C3b inactivator (C3bINA). Addition of IgG anti-E but not IgM anti-E to ETC3b markedly enhanced neutrophil binding and stimulated ingestion of these erythrocytes. There were minimal binding and ingestion of E coated with IgG alone.

Binding of ETC3b to neutrophils was also insufficient to stimulate the release of superoxide anion (O2-), β-glucuronidase, and lysozyme. EIgG, prepared with increasing concentrations of IgG, promoted the release of O2- and granule enzymes in a concentration-dependent fashion, and ingestion was not required. Addition to ETC3b of a small quantity of IgG synergistically enhanced O2- release and degranulation.

These results suggest that the Fc and C3b receptors on neutrophils have separate but complementary functions. C3b serves to promote efficient particle-cell attachment, thereby mediating contact between particle-bound IgG and the neutrophil's Fc receptor. This latter contact triggers ingestion and the intracellular events involved in microbial killing.
Priming and suppression of the intestinal immune response to cholera toxoid/toxin by parenteral toxoid in rats.
Authors: Pierce N F , Koster F T
Published: December 2022

Parenteral immunization of rats with cholera toxoid had both priming and suppressive effects upon the antitoxin response in jejunal lamina propria to locally applied toxoid/toxin. Priming was detected when parenteral toxoid was given i.p. but not i.v. or s.c., was enhanced by Freund's adjuvant, and appeared to reflect enhanced encounter of i.p. antigen with IgA-committed lymphocytes in extra-intestinal mucosa-associated lymphoid tissue. In contrast, suppression followed parenteral toxoid given i.p., i.v., or s.c.; suppression was antigen specific and lasted at least 16 weeks. Parenteral toxoid suppressed both primary and secondary types of mucosal antitoxin responses, ultimately preventing the generation of antitoxin-containing immunoblasts from Peyer's patches. Since suppression followed parenteral immunization by routes that did not provoke mucosal priming, it was, at least in those instances, not simply a regulatory consequence of mucosal priming. These results support the notion that priming and suppression of a specific mucosal immune response are independent effects of parenteral immunization that are probably determined by the distribution of antigen to mucosa-associated and systemic lymphoid tissue, respectively.
Limiting dilution analysis of alloantigen-reactive T lymphocytes. IV. High frequency of cytolytic T lymphocyte precursor cells in MLC blasts separated by velocity sedimentation.
Authors: Maryanski J L , MacDonald H R , Cerottini J C
Published: December 2022

A sensitive limiting dilution microculture system was used to obtain minimal estimates of the frequency of cytolytic T lymphocyte precursor cells (CTL-P) directed against DBA/2 alloantigens, after priming of spleen cells in unidirectional mixed leukocyte cultures (MLC, C57BL/6 anti-DBA/2). The mean CTL-P frequency in day 4 to 5 MLC populations was found to be approximately 50- to 100-fold greater than the frequency in normal spleen, and up to 25% of the cells present in such MLC could be identified operationally as CTL-P. Even higher frequencies (up to 50%) of CTL-P were obtained in a population of large-sized cells separated from day 4 MLC by velocity sedimentation. Furthermore, since a strikingly quantitative correlation was observed between CTL activity and CTL-P frequency in such separated MLC populations, it is likely that mature CTL in MLC are not end cells, but can further proliferate and thus behave operationally as CTL-P.
Viral inhibition of lymphocyte mitogenesis. I. Evidence for the nonspecificity of the effect.
Authors: Wainberg M A , Israel E
Published: December 2022

We have investigated the mechanism(s) whereby virus particles, when co-incubated with lymphocytes, are able to abrogate mitogen- and alloantigen-driven cell proliferation. Our data indicate that this inhibition is entirely nonspecific with regard to its induction; similar results can be obtained by using any of several different types of infectious virus particles or viruses whose infectivity had been destroyed by irradiation with ultraviolet light. Furthermore, we show that plasma membranes vesicles, which approximate viruses in size, and which are derived from normal cells, can also impede lymphocyte proliferative responsiveness. In the mixed lymphocyte culture (MLC) reaction, the inhibitory effect is apparently due to the action of viruseith virus can inhibit the mitogen-induced proliferation of resh syngeneic spleen cells, even if added to the latter at a ratio as low as 1:250. These results suggest that virus particles induce the activation of suppressor lymphocytes which in turn act on those cells which would otherwise be responsive in mitogenesis assays and in the MLC.
The suppression of Epstein-Barr virus infection in vitro occurs after infection but before transformation of the cell.
Authors: Thorley-Lawson D A
Published: December 2022

It has been demonstrated that T cells from humans can suppress the transformation of EBV-infected B cells in vitro. In this paper, studies are presented on the mechanism of the suppression. The T cells do not act against the virus itself nor the infection process. They appear to suppress the outgrowth of B cells after they are infected but before they are transformed. The T cells are much less effective in suppressing the outgrowth of the B cells once the latter have transformed (24th post-infection). The significance of these observations to EBV infection and other forms of T cell-mediated anti-EBV immunity is discussed.
Mechanisms of immune lysis. I. Physiological distinction between target cell death mediated by cytotoxic T lymphocytes and antibody plus complement.
Authors: Russell J H , Masakowski V R , Dobos C B
Published: December 2022

The pattern of progressive release of cytosolic (52Cr) and nuclear (125IUdR) labels from normal or tumor cells being lysed by CTL is strikingly different from the pattern observed when similar cells are lysed by Ab + C or hypotonic shock. We propose that these differences reflect physiologic differences in the lytic events associated with Ab + C- and CTL-mediated cell death.
Separation of a serum-derived tumoricidal factor from a helper factor for plaque-forming cells.
Authors: Männel D N , Farrar J J , Mergenhagen S E
Published: December 2022

C3H/HeN mice administered BCG followed by lipopolysaccharide 14 days later released into their serum a cytotoxic factor for tumor cells and a factor that restored the anti-SRBC plaque-forming cell response of nude mouse spleen cells (helper activity). Gel filtration of serum containing the cytotoxic and the helper activities indicated that both factors exhibited an apparent m.w. of 125,000 to 150,000. The helper activity was also found at lower m.w. (60,000 and 13,000) suggesting the possibility that this factor existed in aggregated forms. Gel filtration of ammonium sulfate (40 to 60% saturation) precipitated serum in a high ionic strength buffer (1.6 M NaCl) resulted in shifts in the apparent m.w. of both factors. The cytotoxic factor now exhibited a m.w. of 55,000. The helper activity eluted with an apparent m.w. of 13,000, and thus was clearly separated from the cytotoxic factor. The helper activity was further shown to co-elute with macrophage-derived lymphocyte activating factor (LAF). This as well as other data represent the first demonstration of in vivo produced LAF.
A monoclonal antibody reactive with the human cytotoxic/suppressor T cell subset previously defined by a heteroantiserum termed TH2.
Authors: Reinherz E L , Kung P C , Goldstein G , Schlossman S F
Published: December 2022

A hybridoma-secreting monoclonal antibody was produced from the spleen cells of a mouse immunized with human thymocytes. This hybridoma antibody, termed OKT5, was reactive by indirect immunofluorescence with 80% of human thymocytes but only 20% of peripheral blood T cells. Moreover, OKT5 was unreactive with normal B cells, null cells, and macrophages at any dilution tested. A similar pattern of reactivity was seen with an equine antiserum to human thymocytes termed anti-TH2. Fluorescence-activated cell sorting demonstrated that the OKT5 antibody reactivity on peripheral T cells was restricted to the majority of the previously defined TH2+ subpopulation. In functional studies, the OKT5+ subset, like the TH2+ subset, proliferated well to the mitogen Con A and to alloantigens, and contained cytotoxic effector cells after sensitization in MLC, and suppressor effector cells after activation with Con A. In addition, like the TH2+ T cell, the OKT+ T cell was virtually unresponsive to soluble antigen. Thus, the OKT5 monoclonal antibody is reactive with the cytotoxic/suppressor T cell subset. OKT5 should provide an important probe to assess the status of suppressor cells in human disease.
Dissociation of differentiation and proliferation in the primary induction of cytolytic T lymphocytes by alloantigens.
Authors: MacDonald H R , Lees R K
Published: December 2022

The requirement for DNA synthesis in the induction of cytolytic T lymphocytes (CTL) by alloantigens has been investigated. C57BL/6 splenic T cells purified by passage on nylon wool columns were stimulated in vitro in mixed leukocyte culture (MLC) and assayed for cytotoxicity against 51Cr-labeled target cells. With this system, CTL activity was detectable after 24 hr of MLC and reached high levels after 48 hr. Addition of cytosine arabinoside (ARA-C) or hydroxyurea to such cultures at concentrations that were sufficient to inhibit DNA synthesis by greater than 98% did not reduce CTL activity measured after 24 hr; however, the increase in activity that occurred between 24 and 48 hr in control cultures was strongly reduced (or abolished) by these drugs. Velocity sedimentation analysis of MLC cells activated for 48 hr in the presence of ARA-C further revealed that CTL precursor lymphocytes had enlarged into medium- to large-sized CTL under these conditions. These studies provide direct evidence that the primary induction of CTL by alloantigens can be dissociated into a differentiation step, which occurs within 24 hr in the absence of DNA synthesis and is accompanied by blast transformation, and a subsequent proliferation.
Antibodies to murine leukemia virus gp70 and p15(E) in sera of BALB/c mice immunized with syngeneic chemically induced sarcomas.
Authors: Klitzman J M , Brown J P , Hellström K E , Hellström I
Published: December 2022

The sera of BALB/c mice immunized with syngeneic methylcholanthrene-induced sarcomas commonly contain antibodies that react with the immunizing tumor line and also with other chemically induced sarcomas. Three lines of evidence suggest that the cross-reactive antibodies are directed against antigenic determinants of gp70 and p15(E), envelope proteins of endogenous murine leukemia virus (MuLV). 1) The antibodies bind only to sarcomas expressing MuLV antigens, 2) they are removed by absorption with purified MuLV antigens, and 3) they precipitate proteins identified as gp70 and p15(E) from 125I-labeled sarcoma cell lysates.
Stability of DNA/anti-DNA complexes. III. Kinetic control of immune complex size.
Authors: Taylor R P , Waller S J , Haden C , Addis D J
Published: December 2022

The Farr assay and sucrose gradient ultracentrifugation were used to study the importance of kinetic factors on the size of antibody/DNA immune complexes prepared from SLE sera and dsDNA. We have found that significantly different numbers and sizes of antibody/DNA immune complexes can be formed by varying the time course of addition of a given amount of DNA to an SLE serum. For example, the introduction of additional DNA to a preincubated antibody-DNA system does not lead to reequilibration of that system with respect to the amount of DNA bound or the size of the complexes formed even after 1 hr at 37 degrees C. The potential implications of these observations with respect to the pathogenesis of SLE is discussed.
IgA1 proteases from Haemophilus influenzae, Streptococcus pneumoniae, Neisseria meningitidis, and Streptococcus sanguis: comparative immunochemical studies.
Authors: Kilian M , Mestecky J , Kulhavy R , Tomana M , Butler W T
Published: December 2022

IgA1 proteases from H. influenzae, N. meningitidis, S. pneumoniae, and S. sanguis were compared with respect to site of cleavage in the IgA1 molecule and EDTA sensitivity. Proteases from S. sanguis and S. pneumoniae cleaved the Pro (227)-Thr (228) bond within the hinge region of the alpha 1 chain and were inhibited by EDTA. H. influenzae IgA1 protease cleaved the Pro (231)-Ser (232) peptide bond. The activity of IgA1 proteases from H. influenzae and N. meningitidis was unaffected by EDTA. Purified and denatured alpha 1 chain was cleaved only in the hinge region. Other component chains of secretory IgA (secretory component, light and J chains) were not susceptible. In addition to IgA1 protease, S. pneumoniae released exo- and endoglycosidases that removed a considerable portion of carbohydrate side chains of IgA1; this activity was absent from crude IgA1 protease preparations of the other three bacterial species. Association in vitro of polymeric IgA1 with SC did not inhibit the degradation of IgA1 proteases. The considerable resistance of secretory IgA to cleavage by IgA1 proteases may be explained in part by the presence of IgA1 protease-neutralizing antibodies in secretory IgA.
Effect of cyclosporin A on human lymphocyte responses in vitro. I. CsA allows for the expression of alloantigen-activated suppressor cells while preferentially inhibiting the induction of cytolytic effector lymphocytes in MLR.
Authors: Hess A D , Tutschka P J
Published: December 2022

The effect of cyclosporin A (CsA) on in vitro human lymphocyte responses was assessed. CsA suppressed in a dose-dependent fashion, the lymphocyte response to stimulation with mitogens and with alloantigens in primary and secondary mixed lymphocyte reactions (MLR). In contrast, the action of cytolytic effector lymphocytes was not affected by this compound. Time-course kinetic studies indicated that the mitogen response was markedly dependent on the time of addition of CsA to the cultures. Inhibition of the lymphocyte response to alloantigens was less dependent upon the time of addition of CsA to the cultures. Preincubation with CsA did not markedly affect the ability of the lymphocytes to respond to mitogens and alloantigens, nor was there an overt cytotoxic action. In addition, CsA was shown to have a differential effect on the activation of cytotoxic and suppressor lymphocyte subpopulations in primary MLR. The induction of cytolytic lymphocytes was markedly suppressed by minimal amounts of CsA, whereas the induction of alloantigen-activated suppressor cells was much less inhibited by this agent. These results suggest that CsA treatment favors the induction of suppressor cell mechanisms as opposed to cytolytic effector cells in primary MLR. This may account for the ability of this drug to establish transplantation tolerance in vivo.
T cell maturation: thymocyte and thymus migrant subpopulations defined with monoclonal antibodies to the antigens Lyt-1, Lyt-2, and ThB1.
Authors: Scollay R , Weissman I L
Published: December 2022

The phenotypes of thymus cells for the antigens Lyt-1, Lyt-2 and ThB have been analyzed by using immunofluorescence techniques. Cells throughout the intrathymic maturation sequence have been tested, including the primitive subcapsular lymphoblasts, all size classes of cortical and medullary thymocytes, and thymus cell emigrants. ThB antigen is not detectable on migrant cells, but all subpopulations in the thymus are subdivided into two categories, bright and dull. Thus, it is possible that the bright and dull phenotypes represent a lineage specific rather than a stage-specific marker, at least inside the thymus. The Lyt-defined thymocyte subclasses Lyt1+2+-(Lyt-1) and Lyt 1+2+ (lyt-1,2) are also both represented in all subpopulations, including subcapsular lymphoblasts. This suggests that they may represent two separate lineages, and that the Lyt-12(3) class is not a precursor of the Lyt-1 class, although the possibility of a very early Lyt-(12(3) cell precursor common to both lines cannot be ruled out.
Idiotypic analysis of anti-GL phi antibodies. I. Identification and strain distribution of GL-1 idiotypes.
Authors: Ju S T , Benacerraf B , Dorf M E
Published: December 2022

We utilized both the inhibition of antigen binding and direct idiotype binding methods to identify a new set of common idiotype determinants on anti-GL antibodies of various mouse strains. Three anti-idiotypic antisera, each prepared against individually purified B10.WB anti-GL phi antibodies, were able to detect antibody-combining site-associated common idiotypic determinants, designated GL-1 idiotype(s), in antisera with GLT-binding activity obtained from all mouse strains except strains bearing Igh-1e allotype. Anti-GL phi antisera obtained from rabbits, guinea pigs, and rats did not express detectable levels of GL-1 idiotypes. Nonresponder mice to GL phi, upon immunization with GL-F gamma G or GL phi-F gamma G produced anti-GL antibodies expressing GL-1 idiotypes. Although the magnitude of the immune response to various GL-containing polymers is controlled by distinct Ir genes, the common GL-related antigenic determinants on these polymers are able to induce anti-GL antibodies with GL-1 idiotypic specificities.
Definitive mapping of the immune response gene(s) for (T,G)-A--L to the I-A subregion.
Authors: Markman M , Dickler H B
Published: December 2022

Mice of strains B10, B10.A, B10.A(4R), B10.A(5R), and the new recombinant inbred strain B10.MBR (H-2 genotype Kb, Ik, Sk. Dq) were immunized with (T,G)-A--L and the primary (total and IgM component), and secondary antibody responses were measured by using radioimmunoassays. B10 and B10.A(5R) were high responders, whereas B10.A, B10.A(4R), and B10.MBR were low responders to (T,G)-A--L. Based on the genotypes of the strains utilized, the immune response gene(s) for (T,G)-A--L is located in the I-A subregion of the H-2 complex. The data are consistent with the concept that Ia antigens are Ir gene products.