bluuebunny/crossref_metadata_2025 huggingface dataset:
Cell-Mediated Cytotoxicity Against Measles Virus in SSPE
Authors: Kreth Hans W. , Wiegand Gerd
Published: December 2022

An analysis of subacute sclerosing panencephalitis (SSPE) lymphocytes was performed in order to find out whether T or K cells were involved in killing of 51Cr-labeled allogeneic measles virus-infected target cells. Lymphocyte donors were three patients with SSPE, 10 measles seropositive controls and 2 children with measles rash. It was found that about 75% of measles-specific cytotoxicity was lost after removal of Fc-receptor-bearing cells by adsorption onto immune complex monolayers. K cell activity (as measured by lysis of 51Cr-labeled-sensitized chicken red blood cells) was reduced to the same extent. After adsorption, the enhancing effect by specific antibody was no longer observed. Unfractionated peripheral lymphoid cells that had been treated with pronase and kept in culture 24 hr were inactive in the cytotoxicity test when compared to freshly isolated cells. However, cytotoxicity could be restored almost completely by the addition of measles antibodies.

The results indicate that measles-specific cytotoxicity by peripheral lymphoid cells from all three groups of donors is mediated by K cells. It is probable that specificity is provided by a small amount of contaminating serum antibody or immune complexes.
Acute Lymphoblastic Leukemia (ALL) Antigens Detected with Antisera to E Rosette-Forming and Non-E Rosette-Forming All Blasts
Authors: Borella Luis , Sen Luisa , Casper James T.
Published: December 2022

Based on the presence or absence of erythrocyte receptors(E) a T cell marker, acute lymphocytic leukemia (ALL), can be divided into E+ALL and E-ALL. We studied cell surface antigens on blasts from 12 children with untreated ALL: eight with E-ALL and four with E+ALL. Heterologous antisera were raised against thymus cells, E+ and E-ALL blasts, approximately absorbed and tested by immunofluorescence and a radiolabeled antibody assay with normal and leukemic lymphoid cells. By both methods, anti-thymus and anti-E+ALL sera reacted with human thymocytes. Specific binding of anti-E+ALL serum to T antigens was indicated by the fact that a single absorption with thymocytes abolished its binding to allogenic thymocytes, and the reactivity of anti-E+ALL serum with thymus, blood and bone marrow lymphocytes was similar to that of anti-thymus serum. After exhaustive absorption with blood leukocytes, anti-E+ALL and E-ALL sera were negative against normal lymphocytes and bone marrow cells from children with ALL in remission. Antithymus and anti-E+ALL sera reacted with blasts from patients with E+ALL, but not with E-ALL. In contrast, anti-E-ALL serum reacted with 40 to 96% of blasts from all children with E-ALL, whereas of the four patients with E+ALL, two were negative and two had the lowest percentage of immunofluorescent cells (10 to 22%). These results were confirmed with the radiolabeled antibody assay. Patients with active E-ALL had cells bearing E-ALL antigen(s) in the peripheral blood and bone marrow, but the number of immunofluorescent cells was lower in blood. Cells reactive with anti-E-ALL sera also were consistently found in sequential bone marrow or blood samples from children with E-ALL who had relapsed and were resistant to chemotherapy. Anti-E-ALL serum did not react with thymus cells, blood lymphocytes, remission bone marrow cells, Raji cells, PWM and PHA-induced blasts and CLL cells bearing mIg (uk). These data suggest that the antigen detected on E-ALL blasts by anti-E-ALL serum is neither a HLA-related nor a cell differentiation antigen.

Thus, by using antiserum to E+ALL blasts, we have confirmed the presence of a T cell-specific antigen(s) on E+ALL cells. This antiserum did not recognize other leukemia-associated antigens common to E+ and E-ALL. We have also demonstrated an antigen(s) which is regularly expressed on E-ALL blasts and is either not detectable or is present in a lower proportion of E+ALL blasts.
IgE Synthesis in Man
Authors: Nagel James , Svec Dianna , Waters Tomi , Fireman Philip
Published: December 2022

Specific IgE and IgG antibodies were measured in 39 adolescent and adult humans preselected to have less than 0.125 antitoxin units (AU) ml of tetanus antitoxin (IgG) irrespective of previous immunization history. Mean antitoxin values (IgG) measured by sheep red cell hemagglutination (SRBC) increased by 2 logs to 1.49 AU/ml sera for tetanus and by 1 log to 0.025 AU/ml sera for diphtheria. Specific tetanus IgE was noted to increase after immunization from a mean 2.0 ± 0.2% counts bound to 3.1 ± 0.6% counts bound, but was not statistically significant. Diphtheria-specific IgE rose significantly (p < 0.05) from a mean 1.5 ± 0.2% counts bound to a mean 4.0 ± 1.2% counts bound in atopic individuals and from a mean 1.2 ± 0.0% counts bound to a mean 2.2 ± 0.5% counts bound in nonatopics. These data document the development of tetanus and diphtheria IgE antibodies after booster immunization.
Coincident Hepatitis B Surface Antigen and Antibodies of Different Subtypes in Human Serum
Authors: Tabor Edward , Gerety Robert J. , Smallwood Linda A. , Barker Lewellys F.
Published: December 2022

Three patients with simultaneously detectable hepatitis B surface antigen (HBsAg) and antibody (anti-HBs) in their sera were studied for subtypes of HBsAg and anti-HBs. In each case anti-HBs was found to be directed to a different subtype than that of the circulating HBsAg, indicating that reinfection (or simultaneous infection) with a second subtype occurred.
Uncommon Gm Phenotypes in Sera from Neuroblastoma Patients
Authors: Morell Andreas , Käser Hans , Scherz Rudolf , Skvaril Frantisek
Published: December 2022

Gm phenotypes and IgG subclass levels were determined in the sera of 68 patients with neuroblastoma. The frequency of the uncommon phenotypes Gm(a+,f+,g-,b+) and Gm(a+,f+,g+,b-) was found to be significantly higher in the neuroblastoma group than in normal blood donors (p < 0.001). IgG subclass concentrations in the nine sera with uncommon Gm phenotypes and in the sera of neuroblastoma patients were inconspicuous.
Complete Humoral Immunosuppression of Mice by Rabbit Anti-μ Antibodies Passing the Murine Placenta
Authors: Manning Dean D.
Published: December 2022

Female BALB/c mice were immunosuppressed with a series of neonatally initiated rabbit anti-μ serum injections, which eliminated serum IgM and greatly delayed production of antibodies against normal rabbit serum (anti-NRS). Females thus prepared maintained circulating anti-μ levels for several months. Study of the progeny from pregnancies completed during this maintenance period revealed that rabbit anti-μ antibodies readily cross the murine placenta but are not passed in murine colostrum at levels detectable by the technique used. Anti-NRS antibodies actively produced in NRS-injected control females do cross the placenta, but do so only irregularly and poorly; these antibodies may, however, be detected consistently at relatively low levels in colostrum. Suppression of humoral immunoglobulin synthesis in most mice prenatally exposed to anti-μ antibodies by transplacental passage appeared complete, even including loss of the remnant IgG levels which are consistently seen in mice first exposed to anti-μ at birth. The appearance of serum IgG and anti-NRS antibodies along with the complete absence of serum IgM in mice recovering from suppression suggests that active IgM synthesis and secretion may not be a prerequisite for the IgM to IgG “switch.” Immune recovery occurred even in completely immunosuppressed mice after anti-μ injections were discontinued; the mechanism of recovery is not certain.
Fractionation of Lymphocytes Involved in the Generation of Cell-Mediated Cytotoxicity over Insolubilized Conjugated Histamine Columns
Authors: Shearer Gene M. , Simpson Elizabeth , Weinstein Yacob , Melmon Kenneth L.
Published: December 2022

Spleen cells from normal BALB/c mice were cultured in vitro with irradiated C57BL/6 stimulating cells. Five days later the T cell-mediated cytotoxic activity of the effector cells was assessed with a 51Cr-release assay that used H-2bEL-4 tumor cells as targets. Before the BALB/c responding lymphocytes were sensitized they were fractionated by passing the spleen cells over insolubilized histamine rabbit serum albumin Sepharose columns (H-RSA-S) or over rabbit serum albumin Sepharose (RSA-S) control columns. Fractionation of cells over the H-RSA-S columns depleted or significantly reduced the cytotoxic potential of the unretained cells. All cytotoxic potential was recovered when the cells that adhered to the H-RSA-S were eluted from the columns. In contrast, no effect on responsiveness was detected after the cells had been fractionated over the control column. The loss of response potential by the cells that did not adhere to H-RSA-S could not be accounted for by removal of macrophages nor by the concentration of cells with suppressor activity in the effluent. These cell fractionation studies raise the possibility but do not prove that cytotoxic precursor cells may express amine receptors that could be responsible for their retention by insolubilized histamine columns.
Selective Proteolysis of the J Chain Component in Human Polymeric Immunoglobulin
Authors: Koshland Marian Elliott , Chapuis Rose Marie , Recht Bernard , Brown John Clifford
Published: December 2022

To clarify the losses that have been observed in the J chain portion of human IgM and IgA, were carried out studies on the enzymatic susceptibility of the J polypeptide. When Waldenström macroglobulins and myeloma IgA polymers were subjected to limited proteolysis with various endopeptidases, only subtilisin was found to attack the J chain component. The pattern of cleavage was a function of the polymer species. The J chain in IgM was highly susceptible to digestion, quantitative cleavage being achieved at very low enzyme to IgM ratios and without significant changes in the remaining pentamer structure. Analyses of the digestion products showed that the initial cleavage occurred at an exposed region midway in the J sequence and was followed by extensive degradation of the carboxy-terminal segment. These findings indicated that the observed loss of the IgM J component can be explained by the inadvertent introduction of subtilisin in vitro or by the attack of in vivo enzymes with a specificity similar to subtilisin. In contrast, the IgA J chain was found to be much more resistant to subtilisin proteolysis; its cleavage required higher enzyme concentrations and was accompanied by significant degradation of the α-chains. Thus, it appears unlikely that the IgA J polypeptide is degraded by either in vitro or in vivo enzymes unless its accessibility is first enhanced by changes in the IgA Fc structure.
Relationship between Trinitrophenyl and H-2 Antigens on Trinitrophenyl-Modified Spleen Cells
Authors: Forman James , Vitetta Ellen S. , Hart David A. , Klein Jan
Published: December 2022

Spleen cells were treated with TNBS in order to determine if cell surface H-2 antigens are derivatized with TNP. By labeling the cell membrane of the TNP-modified cells with 125I, followed by detergent lysis and immune precipitation with anti-TNP, it was determined that no H-2 antigenic activity remained in the supernatant. Further, by the use of an antibody-induced antigen redistribution assay it was found that previous exposure of TNP-modified cells to anti-TNP in the absence of complement rendered these cells resistant to lysis by anti-H-2 in the presence of complement. Together these data indicate that at the concentration of TNBS used for modification, H-2 antigens are derivatized with TNP. However, in addition to H-2, other proteins including immunoglobulin were also derivatized with TNP.

Anti-TNP cytotoxic effector cells were blocked from their cytotoxic activity by anti-TNP antiserum. These data indicate that TNP directly couples to H-2 antigens on the cell surface of TNP-modified cells and that TNP is associated with the antigenic determinant that the cytotoxic T cell recognizes.
Relationship between Trinitrophenyl and H-2 Antigens on Trinitrophenyl-Modified Spleen Cells
Authors: Forman James , Vitetta Ellen S. , Hart David A.
Published: December 2022

Spleen cells were modified with varying concentrations of trinitrobenzene sulfonic acid and then assayed for both their ability to stimulate syngeneic spleen cells into displaying a cytotoxic effect against TNP-modified target cells and for the extent of TNP derivatization of H-2 antigens. It was found that there was a direct correlation between the extent of derivatization of H-2 antigens and the ability of such derivatized cells to act as stimulator cells in the TNP-CML assay. Thus, these data lend support to the altered self or interaction antigen hypothesis as the explanation for the H-2 gene restriction of syngeneic CML.

Target cells were also modified with TNBS at varying concentrations to determine the optimal concentration required to permit lysis in the CML assay. The results of these experiments indicate that similar concentration ranges of TNBS are required to create antigenic determinants on the target cells as well as immunogenic determinants on the stimulator cells that can be recognized by cytotoxic T cells.
Regulation of Antibody Response in Different Immunoglobulin Classes
Authors: Kimoto Masao , Kishimoto Tadamitsu , Noguchi Sanai , Watanabe Takeshi , Yamamura Yuichi
Published: December 2022

In vitro induction of anti-DNP IgE as well as IgG1, IgG2a antibody responses was shown in murine spleen cell culture. Spleen cells primed three times with 1 μg of DNP-OA or DNP-Asc produced significant amounts of anti-DNP IgE as well as IgG antibodies by the in vitro stimulation with DNP-OA or DNP-Asc, respectively. Collaboration between DNP-primed B cells and carrier-primed T cells was required for the induction of both IgE and IgG antibodies with DNP-coupled T-dependent antigen. Carrier-specific T cells induced with a low dose of Asc (0.01 μg) showed helper function only on IgE antibody response, whereas T cells primed with a higher dose of Asc (10 μg) cooperated only with IgG-B cells. T cells primed with Asc in CFA showed helper function mainly on IgG antibody response but not on IgE antibody response. The result indicated the presence of a distinct population of T helper cells for IgE and IgG antibody responses. T-independent antigen (DNP-Ficoll) induced both anti-DNP IgE and IgG antibody responses in DNP-primed spleen cell population without the requirement of the collaboration of helper T cells.
Specificities of Killing by Cytotoxic Lymphocytes Generated in Vivo and in Vitro to Syngeneic SV40 Transformed Cells
Authors: Gooding Linda R.
Published: December 2022

Cell-mediated immunity to SV40-transformed C3H and C3H·SW cell lines was measured by using both 51Cr and 125IUdR release assays. Killing by cytotoxic cells generated on in vitro sensitization of immune spleen cells with syngeneic SV40 cells by either assay is specific for syngeneic SV40 transformants. Cytolysis mediated by in vitro sensitized cells is ablated by treatment of the effector cells with anti-θ serum and complement. Intraperitoneal immunization with syngeneic SV40 cells yields two distinct killer-cell populations in the peritoneal exudate when assayed by 125IUdR release. The first, nylon wool nonadherent and sensitive to anti-θ and complement, is indistinguishable from the killers generated in vitro. The second population, present in larger numbers and more efficient on a per-cell basis in killing of SV40 targets than the first, is nylon adherent and is not removed by treatment with anti-θ and complement. This second population will kill any SV40 transformed target, whether syngeneic or allogeneic. The possible roles of T cell and non-T cell effectors in rejection of syngeneic SV40 tumors are discussed.
Characterization of the Blastogenic and Cytotoxic Responses of Normal Mice to Ecotropic C-Type Viral gp71
Authors: Lee John C. , Ihle James N.
Published: December 2022

Spleen cells from normal (B6C3)F1 mice demonstrated natural cytotoxic reactivity mediated by a “null” cell population which, in part, had immunologic specificity for the major envelope glycoprotein (gp71) of the endogenous ecotropic murine leukemia virus. In contrast, the cytotoxic reactivity reflected in spleen cells of NIH Swiss nude mice apparently had no immunologic specificity. A significant level of blastogenic response could be generated in vitro by using normal (B6C3)F1 and AKR spleen cells in the presence of gp71. This reactivity was highly type specific. Moreover, using normal spleen cells from (B6C3)F1, both secondary blastogenic responses and cytotoxic T cell responses could be induced in vitro with purified soluble viral gp71. These findings extend further our previous studies on the existence of a natural immune response in normal mice to their endogenous MuLV by providing in vitro evidence for the expression of cell-mediated response in addition to the humoral response and that at least two effector cell types are operable in the cell-mediated phase.
Recovery of Soluble Sheep Erythrocyte Receptor from the T Lymphocyte Surface by Proteolytic Cleavage
Authors: Chisari Francis V. , Gealy W. James , Edgington Thomas S.
Published: December 2022

Proteolytic digestion of the human T lymphoblastoid cell line (Molt-4) and of peripheral blood lymphocytes by trypsin, chymotrypsin, and pronase results in a progressive, time- and dose-dependent diminution of T lymphocyte — sheep red blood cell (SRBC) rosette formation, whereas thrombin, plasmin, collagenase, DNAse, and phospholipase have no effect. Complete abrogation of SRBC binding is achieved when lymphocytes (1 × 108/ml) are incubated with either trypsin or chymotrypsin at 10 μg/ml for 30 min, and τ;50% abrogation is observed between 3 to 10 min. Preincubation of SRBC with the 10 min and 20 min lymphocyte digest supernatants inhibited their subsequent binding by normal T lymphocytes by as much as 64%. Thirty-minute digests were less inhibitory. Equivalent digests from several human B lumphoblastoid cell lines and from a non-rosetting clone of Molt-4 cells were not inhibitory. Polyacrylamide gel electrophoresis followed by elution of serial gel slices revealed four distinct inhibitory bands (I–IV) in the 20-min digest supernatant whereas only bands I–III and band IV were present in the 10-min and 30-min digest supernatants, respectively, suggesting progressive proteolysis of a distinct receptor. These experiments indicate that the binding of SRBC by human T lymphocytes represents a receptor-ligand interaction rather than a nonspecific electrical charge phenomenon and that the receptor is a discrete molecular species which can be isolated from the surface of T but not B lymphocytes by limited enzymatic proteolysis.
Immunologic Release of Heparin from Purified Rat Peritoneal Mast Cells
Authors: Yurt Roger W. , Leid R. Wesley , Spragg Jocelyn , Austen K. Frank
Published: December 2022

High m.w. [35S]heparin, labeled in vivo or in vitro, was released from purified rat mast cells by challenge with rabbit anti-rat F(ab')2, guinea pig anti-rat IgE, or calcium ionophore. The released and the residual heparin were isolated by Dowex 1 chromatography and were of comparable size by Sepharose 4B gel filtration. The majority of the released heparin was found by differential centrifugation to be granule-associated. Net percentage of mast cell heparin release, quantitated by metachromasia after isolation on Dowex 1 chromatography, correlated in a linear fashion with net percentage of histamine release, with heparin exhibiting a threshold requirement for onset of release. The correlation of histamine and high m.w. heparin release provides chemical support for the conclusion of others from ultrastructural studies that mast cell activation by immunologic means or by the calcium ionophore results in secretion of the whole granule.
B Lymphocyte Colony-Forming Cells in the SJL/J Mouse Thymus
Authors: Claesson Mogens H. , Metcalf Donald
Published: December 2022

Mercaptoethanol-induced B lymphocyte cloning in semisolid agar has been used to study lymphocyte colony formation by cells from the SJL/J mouse thymus. From the 3rd month of life, the SJL thymus develops an increasing frequency of cells forming B lymphocyte colonies in agar. The peak frequency in 6- to 12-month-old mice was one colony per 1000 to 2000 cultured thymus cells. In contrast, 10 to 100 times lower frequencies were found in the thymus of five other inbred mouse strains. The rise in B lymphocyte colony-forming cells correlated well with the age-related rise in Ig-positive cells and approximately 50% of the colony cells reacted with anti-μ-serum indicating the B lymphocyte nature of the colony cells. Colony-forming cells from the thymus showed higher sensitivity than colony-forming spleen cells to cortisol and irradiation.

Cell transfer experiments and thymus grafting suggested that the increased frequency of colony-forming cells in the thymus is caused by development of special thymus-seeking B lymphocytes in ageing SJL/J mice. Finally, B lymphocyte colony-forming cells were found to be more frequent in the thymus, spleen, and lymph nodes from healthy aged mice than in lymphoid organs from mice with spontaneous reticulum cell tumors.
Studies of H-2 Restriction in Cell-Mediated Cytotoxicity and Transplantation Immunity to Leukemia-Associated Antigens
Authors: Ting Chou-Chik , Law Lloyd W.
Published: December 2022

H-2 dependency of T cell-mediated cytotoxicity and transplantation immunity to leukemia-associated antigens has been investigated. Through the use of a 20-hr 125IUdR release assay, it was found that the induction of T cell-mediated cytotoxicity against Friend virus-induced leukemias of different H-2 haplotype origins could be produced by immunization with both syngeneic and allogeneic tumor cells; the effector cells that were generated by syngeneic immunization could also provide effective killing of allogeneic tumor cells, although the killing of allogeneic targets might require a longer incubation time (20 to 40 hr). Furthermore, in vivo transplantation immunity against Friend virus-induced leukemias also was induced by immunization with both syngeneic and allogeneic tumors and syngeneic immunization could induce specific protection against the challenge with allogeneic tumor in x-irradiated hosts. These findings clearly indicate that, both at the sensitizing phase and effector phase of the immune response, there is no strict H-2 dependency for T cell-mediated cytotoxicity or in in vivo transplantation imunity to leukemia-associated antigens.
Experimental Allergic Encephalomyelitis in the Lewis Rat: Further Delineation of Active Sites in Guinea Pig and Bovine Myelin Basic Proteins
Authors: Martenson Russell E. , Nomura Kohji , Levine Seymour , Sowinski Richard
Published: December 2022

Highly encephalitogenic peptide (37–88), derived from the guinea pig myelin basic protein by peptic digestion, was treated chemically to destroy its tyrosyl and histidyl residues and enzymatically to remove its C-terminal sequence Val-His-Phe. Neither of the modifications resulted in loss of activity in Lewis rats. The encephalitogenic region within peptide (37–88) was located by examination of derivative peptides obtained by selective proteolytic cleavage. The results showed that peptide (61–88), like peptide (43–88), was fully active at the level of 0.02 nmole whereas peptides (72–88) and (72–84) were fully active at levels of 0.5 and 2.5 nmole, respectively. In contrast, peptides (43–71) and (75–88) were completely inactive. These results demonstrated that the undecapeptide Gln-Lys-Ser-Gln-Arg-Ser-Gln-Asp-Glu-Asn-Pro (residues 72–84), although not as encephalitogenic as peptides (43–88) or (61–88), does contain the elements essential for the induction of disease. At the levels tested (10.8 and 2.2 nmole) only peptides (43–88) and (61–88) were capable of inhibiting the induction of disease by passively transferred lymph node cells; this inhibition, however, was less than that achieved by the intact guinea pig basic protein. Further studies on the encephalitogenicity of the bovine basic protein in Lewis rats demonstrated that the active site in the C-terminal half of this protein is present in its entirety within residues 89 to 115.
Recall of Specific Suppression: Co-Dominance of Suppression after Primary or Secondary Antigen Stimulation
Authors: Eardley Diane D. , Sercarz Eli E.
Published: December 2022

Mouse spleen cells primed in vivo with β-galactosidase (GZ), and cultured in vitro with trinitrophenyl β-galactosidase (TNP-GZ), make a very poor anti-TNP response compared to cultures of normal spleen cells. The lack of response is caused by active suppression as indicated by cell mixture experiments. At 2-months following priming, while the helper effect predominates, suppression can be recalled by rechallenge of the mice with GZ. Both primary suppression (after a single injection) and secondary suppression (after two injections) reduce the response of normal or carrier-primed helper cells at equivalent cell ratios in mixture experiments. Both kinds of suppression are carrier specific, selectively suppress high avidity antibody production, and require cell division to suppress the response of normal cells. The only difference between primary and secondary suppression is that primary suppression is obliterated in cultures which are not challenged with antigen until 24 hr after cultures are established. Secondary suppression is still observed when cultures receive TNP-GZ at 24 hr. Suppression appears to be an early component of both primary and secondary responses. The regulatory function of such suppression may be the delay of antibody secretion.
Immunity to Sporozoite-Induced Malaria Infection in Mice
Authors: Chen David H. , Tigelaar Robert E. , Weinbaum Frederic I.
Published: December 2022

The cellular basis of immunity to sporozoites was investigated by examining the effect of immunization of T and B cell-deficient C57BL/6N × BALB/c AnN F1 (BLCF1) mice compared to immunocompetent controls. Immunization of T cell-deficient (ATX-BM-ATS) BLCF1 mice with x-irradiated sporozoites did not result in the generation of protective immunity. The same immunization protocols protected all immunocompetent controls. In contrast, B cell-deficient (μ-suppressed) BLCF1 mice were protected by immunization in the majority of cases. The absence of detectable serum circumsporozoite precipitins or sporozoite neutralizing activity in the μ-suppressed mice that resisted a sporozoite challenge suggests a minor role for these humoral factors in protection. These data demonstrate a preeminent role for T cells in the induction of protective immunity in BLCF1 mice against a P. berghei sporozoite infection.
Priming for a Cytotoxic Response to Minor Histocompatibility Antigens: Antigen Specificity and Failure to Demonstrate a Carrier Effect
Authors: Bevan Michael J.
Published: December 2022

Spleen cells from normal mice do not give a detectable in vitro cytotoxic T cell (CTL) response to minor H antigens. Spleen cells from animals primed in vivo with minor H antigens give a strong CTL response when boosted in culture with the appropriate stimulating cells. Here I have studied the requirements for priming a CTL response to minor H antigens. It is shown that priming is just as antigen specific as is cytolytic effector function. That is, the priming cells have to carry the same minor antigens as the challenge cells. Inducing a graft-vs-host reaction in vivo does not nonspecifically allow spleen cells to respond to minor H antigens in vitro.

Using minor H congenic mice (congenic for H-Y and/or H-7) I have also tried, and failed, to demonstrate a carrier effect in priming. Female mice primed to H-Y were challenged in culture with cells bearing H-Y and H-7 antigens in the hope that a helper response to H-Y would augment a CTL response to H-7. This did not happen, however. Such primed and boosted cells gave a strong secondary CTL response to H-Y but none to H-7. It is concluded that in order to prime for a detectable in vitro response to minor antigens it is necessary to expose the CTL precursors to antigen in vivo. This either expands the size of the pool of precursors by cell division or changes them in some qualitative way.
Long-Term Effects of in Vitro Sensitization on Immune Reactivity of Lymphocytes: Generation of Suppressor and Helper T Cells
Authors: Chan Eva L. , Kan Patricia B. , Mishell Robert I.
Published: December 2022

Mouse spleen cells were cultured for 5 days with or without HRBC. Cultured cells were “parked” in irradiated syngeneic recipients for 3 weeks and then tested for their immunologic reactivity in vitro. We found that spleen cells from recipients of HRBC-sensitized cells (S) as well as spleen cells from recipients of control unsensitized cells (U) possessed radiosensitive suppressor and radioresistant helper activities. Suppressor activity was observed by the capacity of unirradiated S and U spleen cells to inhibit the in vitro generation of IgM and IgG PFC by spleen cells primed in vivo to HRBC or to LacKLH. Helper activity was shown by the capacity of the irradiated S and U cells to restore IgM and IgG PFC responses of in vivo primed, T-depleted spleen cells to HRBC, LacHRBC, and LacCRBC. Both suppressor and helper activities were mediated by T cells. The possibilities that immunologically specific or nonspecific mechanisms account for these phenomena are discussed.
The Allogeneic Effect on Tumor Growth
Authors: Osborne David P. , Katz David H.
Published: December 2022

The growth of an ascitic murine plasmacytoma, MOPC 315, can be retarded in CAF1 hybrid host mice by the i.p. injection of donor lymphoid cells. The graft-vs-host reaction can be established by a variety of donor cells, including parental BALB/c and A/J and congenic inbred B10.D2 which share the major histocompatibility locus with BALB/c(H-2d). Optimal results are consistently obtained when parental BALB/c spleen cells are injected before tumor inoculation, and a second dose of donor spleen cells injected 1 week later. This allogeneic effect on tumor growth is manifested by delayed appearance of the tumor and prolonged host survival. Pathologic studies on the ascites tumor indicate that the allogeneic effect suppresses the initial appearance and early growth of the plasmacytoma. However, once established, MOPC 315 grows rapidly and fatally in both control mice and recipients of donor lymphoid cells. Further, a subcutaneous implant of MOPC 315 is suppressed by an allogeneic effect established either i.v. with BALB/c spleen cells before tumor inoculation or by BALB/c spleen cells administered subcutaneously at the time of MOPC 315 implant. Thirty percent of mice treated by i.v. or subcutaneous donor lymphoid cells were tumor free at 150 days after tumor inoculation.
Are Actinomyces Viscosus Antigens B Cell Mitogens?
Authors: Burckhardt Johann J. , Guggenheim Bernhard , Hefti Arthur
Published: December 2022

An extracellular heteroglycan (ECHG) and a sonicated cell supernatant (SCS) of Actinomyces viscosus Ny 1 induced strong lymphocyte proliferation. This was shown with spleen and thoracic duct cells from germfree rats and confirmed with cells from conventional “nude” mouse spleens. Spleen cells developed direct plaque-forming cells against densely coupled TNP-SRBC. The mitogenic property of ECHG was diminished considerably after mild alkaline hydrolysis for lymphocytes from rat spleens and was totally abolished for cells from “nude” mouse spleens. These results suggest that ECHG and SCS have B cell mitogenicity.
Suppression of Natural Killer Cell Activity with Radioactive Strontium: Effector Cells Are Marrow Dependent
Authors: Haller Otto , Wigzell Hans
Published: December 2022

NK cells with lytic capacity for Moloney leukemia virus-induced lymphomas have previously been found to occur spontaneously in spleens from nonimmune adult mice. Here, 89Sr-treatment is shown to suppress NK cell function in adult mice without similarly affecting other cell-mediated immune reactions. Thus, selective 89Sr-sensitivity distinguishes NK cells from other killer cell types. The present results indicate that in vivo a functional bone marrow is needed for generation and maintenance of NK activity.
Effects of Neonatal Thymectomy and Splenectomy on Survival and Regulation of Autoantibody Formation in NZB/NZW F1 Mice
Authors: Roubinian Jirayr R. , Papoian Ruben , Talal Norman
Published: December 2022

NZW F1 (B/W) mice were subjected to sham surgery or neonatal thymectomy and/or splenectomy and studied for immunoglobulin class of antibodies to double-stranded DNA and polyadenylic acid (Poly A) at 4 to 13 months of age. These antibodies occur spontaneously during the course of autoimmune disease in B/W mice. Sera were fractionated by sucrose density gradient ultracentrifugation and assayed for antibodies by a filter radioimmunoassay method. IgM was recovered in the 19S region and IgG in the 7S region as demonstrated by immunodiffusion.

In sham-operated controls, at all ages studied, anti-DNA antibodies were both IgM and IgG, with the former predominating in males, and the latter in females. In both sexes, anti-Poly A antibodies were primarily IgM in young mice. There was a sequential switch from IgM to either enhanced or new IgG production in the following sequence: female anti-DNA and anti-Poly A (6 months), male anti-DNA (9 months), and male anti-Poly A (11 months). Both thymectomy and splenectomy caused earlier death in male mice, whereas females lived significantly longer after thymectomy. Neonatal thymectomy in males caused a premature switch from IgM to IgG antibodies to DNA, but it had a transient effect in females. Thymectomy almost completely prevented the late switch to IgG antibodies to Poly A in both sexes. By contrast, splenectomy promoted the formation of IgG antibodies to Poly A in male mice.

These results suggest that the newborn B/W thymus and spleen contain regulatory cells and/or factors exerting different controlling influences on spontaneous antibodies to DNA and Poly A. Male B/W mice appear to be under the regulatory influence of suppressor cells, whereas the predominant regulation in female B/W mice appears to be a helper effect.
Studies on Antigen-Induced Arthritis in Mice
Authors: Brackertz Dieter , Mitchell Graham F. , Vadas Mathew A. , Mackay Ian R. , Miller Jacques F. A. P.
Published: December 2022

Antigen-induced arthritis was developed in mice as a model of human rheumatoid arthritis by using methylated bovine serum albumin (mBSA) as antigen. It was found that most strains were susceptible, whereas CBA mice were resistant. We therefore investigated the humoral and cell-mediated immune responses to mBSA in resistant mice (CBA) and susceptible mice (exemplified by C57BL) to determine whether these were associated with susceptibility to arthritis.

The resistant strain (CBA) differed from the susceptible strains in the following respects. First, there was a lower humoral immune response to mBSA as measured by passive hemagglutination, but this could be overcome by a larger immunogenic dose. Secondly, there were differences in response to low doses of DNP-mBSA after mBSA carrier preimmunization. Thirdly, there were striking differences in delayed-type hypersensitivity (DTH) to mBSA as determined by a radioisotopic assay in vivo; the response of CBA mice occurred early, at 5 days, declined quickly, and was weaker, whereas that of C57BL mice developed later and was long sustained. Genetic studies of the DTH response with hybrids and backcrosses showed an oligogenic control of immune responsiveness, with one gene being linked to the H-2b allele of the susceptible C57BL mice, and another being independent of the H-2 complex.

Our findings indicate that in mice, susceptibility to antigen-induced arthritis with mBSA correlates with a higher responder state to this antigen, and that T cells are the major if not the only determinant of the high responder state.
Studies on Antigen-Induced Arthritis in Mice
Authors: Brackertz Dieter , Mitchell Graham F. , Vadas Mathew A. , Mackay Ian R.
Published: December 2022

Antigen-induced arthritis in mice occurs after immunization and subsequent intraarticular challenge with methylated bovine serum albumin (mBSA). In adoptive transfer experiments, susceptible C57BL mice and resistant CBA mice were compared in their capacity to express delayed-type hypersensitivity (DTH) by ear assay, and to express arthritis. The expression of DTH could be transferred incrementally by lymphoid cells in C57BL mice, but not in CBA mice. Both immune lymphoid cells and, to a much lesser extent, serum transferred the capacity to develop arthritis in C57BL mice. The reactivity of transferred cells was abolished by anti-Thy-1 but enhanced by enrichment for T cells with anti-immunoglobulin columns. If this model disease can be equated with human rheumatoid synovitis, the lesions in the human disease would be an expression of a T cell-dependent activity
Antigen Modulation of Antibody Forming Cells: The Relationship between Direct Plaque Size, Antibody Secretion Rate and Antibody Affinity
Authors: Perelson Alan S. , Goldstein Byron
Published: December 2022

Using the mathematical theory of direct plaque growth, we have analyzed the expected variation of plaque size with IgM affinity and secretion rate. We use the theory to comment on recent effector cell blockade experiments and show how the theory can be used to determine the change in the secretion rate of a single antibody-forming cell subjected to blockade by a multivalent antigen. We also argue, using the mathematical theory, that under the usual experimental conditions employed in the plaque assay, cells that produce low affinity IgM antibodies will give rise to smaller plaques than cells that produce high affinity IgM antibodies.
The in Vitro Effects of Histamine and Metiamide on Neutrophil Motility and Their Relationship to Intracellular Cyclic Nucleotide Levels
Authors: Anderson Ronald , Glover Annemarie , Rabson Arthur R.
Published: December 2022

Histamine at concentrations of 1 × 10-5 M to 5 × 10-5 M consistently increased neutrophil movement as measured in Boyden chambers. This effect was entirely caused by stimulation of chemokinesis (stimulated random migration) and true chemotaxis was inhibited by these concentrations. This inhibition of chemotaxis could be abolished by pretreatment with metiamide, an H-2 receptor antagonist, and levamisole, but not by diphenylhydramine, an H-1 receptor antagonist. Metiamide at similar concentrations produced a mild stimulation of chemokinesis but had no effect on true chemotaxis. The histamine effects on neutrophil motility were associated with increased levels of intracellular cAMP whereas cGMP levels were unaffected. Agents known to elevate intracellular cAMP levels produced effects on neutrophil motility similar to those of histamine. It is suggested that histamined exerts a 2-fold effect on neutrophil motility mediated via an H-2 receptor site and associated with elevated levels of cAMP.
Distinctive Functional Properties of Human Blood L Lymphocytes: A Comparison with T Lymphocytes, B Lymphocytes, and Monocytes
Authors: Horwitz David A. , Garrett Martha Anne
Published: December 2022

Human blood lymphocytes with high affinity Fc receptors have been operationally named L lymphocytes because of membrane-labile IgG markers. L lymphocytes lack membrane-incorporated immunoglobulin and do not form rosettes with sheep red blood cells coated with IgM antibody and mouse complement. These lymphocytes are capable of binding IgG in normal human serum at 4°C and will form rosettes with human lymphocytes coated with Ripley IgG. In this study, functional in vitro properties of isolated L lymphocytes were compared with T lymphocytes, B lymphocytes, and monocytes. To obtain these mononuclear populations, first, plastic adherent monocytes were harvested. T lymphocytes were then isolated by centrifugation of E rosette-forming cells, and other rosetting techniques were employed to isolate L and B lymphocytes by negative selection.

The functional properties of L lymphocytes were completely unlike those of T cells, B cells, or monocytes. L lymphocytes did not proliferate in response to mitogens, soluble antigens, or cell surface antigens. Moreover, this population could not replace monocytes in helping T lymphocytes respond to concanavalin A and pokeweed mitogen. Once T cells were supplemented with monocytes, however, the addition of L lymphocytes to the culture greatly enhanced the T lymphocytes proliferative response to phytohemagglutinin, concanavalin A, purified protein derivative (PPD), and streptokinase/streptodornase.

L lymphocytes were not a subset of B cells. They did not spontaneously develop surface Ig in culture, and pokeweed mitogen could not induce them to transform and generate cytoplasmic Ig detectable by immunofluorescence. Mixtures of B cells and T cells responded to pokeweed mitogen better than do T cells alone. In contrast, enhanced reactivity with L and T cell combinations was not observed. Another sharp difference between these two populations was the stimulator capacity of each in mixed lymphocyte culture. When B and L lymphocytes were carefully monocyte-depleted, only B cells were effective stimulators of autologous and allogeneic lymphocytes.

In comparison with T cells, B cells, and monocytes, L lymphocytes were the only effective killers of human blood lymphocytes sensitized with IgG. L lymphocytes, then, have cytotoxic potential, but cannot proliferate in response to various stimulants or become antibody-producing cells. These findings suggest that L lymphocytes comprise a third lymphocyte population.
Further Studies on the Identification of the Subcomponents of the First Component of Complement After Affinity Chromatography of Human Serum on IgG-Sepharose
Authors: Taylor P. A. , Fink S. , Bing D. H. , Painter R. H.
Published: December 2022

Affinity chromatography of serum on IgG covalently linked to Sepharose results in the retention of the proteins of the first component of complement (C1). A fraction called pool II is eluted from this column with 0.025 M EDTA and has previously been shown to contain C1s and a novel protein believed to be part of the C1 complex and called C1t. C1r has now been located in pool II and these three proteins were purified by DEAE cellulose chromatography. C1r and C1s were recovered in the proenzyme form and their identity was established by SDS polyacrylamide gel electrophoresis before and after reduction and alkylation, and on the basis of their esterolytic activities toward different substrates. The properties of C1r from pool II are contrasted with those of the protein recovered from the pool III eluate of the affinity column and previously thought to be C1r.
Sequential Proliferation Induced in Human Peripheral Blood Lymphocytes by Mitogen
Authors: Winger Larry A. , Nowell Peter C. , Daniele Ronald P.
Published: December 2022

Human peripheral blood cells from cultures stimulated by phytohemagglutinin (PHA) for 3 days suppressed continued proliferation when added to feeder layer cultures of autologous lymphocytes previously stimulated with mitogen. Proliferation 1 day after the addition of inhibitory cells was reduced 50 to 60% as measured by tritiated thymidine (3HTdR) uptake and mitotic index. It is suggested that a similar suppressive phenomenon may be acting in most standard mitogen-stimulated cultures to limit cell division after only a few days of culture.

Suppression of continued lymphocyte proliferation in the feeder layer cultures by cells from PHA-stimulated cultures was not due to cell crowding, carry-over of mitogen, or nutrient depletion; nor was it an artifact of thymidine incorporation. Apparent suppression of proliferation by an extract of these cells, however, was artifactual; the extract inhibited 3HTdR uptake but did not reduce mitotic activity.

It is not known whether the inhibition exerted by the PHA-activated cells is mediated through a discrete subpopulation of “suppressor cells” or represents a regulatory function of any mitotic or post-mitotic cell, or both.
Pathogenesis of Experimental Allergic Orchitis
Authors: Tung Kenneth S. K. , Leong Calvin , McCarty Teresita
Published: December 2022

In experimental allergic orchitis (EAO), a lesion characterized by mononuclear invasion of seminiferous tubules can be adoptively transferred within 1 to 4 days by testicular injection of peritoneal exudate cells (PEC) from syngeneic strain 13 guinea pigs (GP) immunized with homologous testicular antigens in complete Freund's adjuvant (CFA). This study examined the role of T lymphocytes, macrophages, and polymorphonuclear neutrophils (PMN) in the adoptive transfer. Guinea pig PEC contained 7% T lymphocytes, rare B lymphocytes, and over 90% of macrophages and PMN. After T lymphocytes were depleted by rabbit erythrocyte (E) rosette and Hypaque-Ficoll gradient centrifugation, cell preparations that contained 73% of original macrophages and 15% original T lymphocytes were obtained, and these cells did not transfer EAO (0 of 18 testes). In contrast, cell preparations enriched in T lymphocytes by nylon wool column or E rosette contained 1.5% of the original macrophages and 59% of the original T lymphocytes transferred EAO to 70% of the testes, starting at 1.5 × 106 T lymphocytes per testis. The number of T lymphocytes correlated with the incidence of adoptive transfer; the correlation existed regardless of the number of macrophages or PMN present. Finally, EAO was adoptively transferred to recipients that had total-body irradiation. The results indicate that (a) T lymphocytes are capable of transferring lesions of EAO, (b) in the transfer, the T lymphocytes did not function as helper T cells, since the transfer need not involve participation of host lymphoid cells, and (c) by inference, testis antigen-reactive T lymphocytes exist.
Preferential Assignment of Allotype a1 Globulin for the Production of Early IgM Anti-Para-Azobenzenearsonate Antibodies in Heterozygous a1a3 Rabbits
Authors: Chien Chau-Chun , Ingraham Joseph S.
Published: December 2022

Rabbits of allotype a1a3 were injected on days 0, 2, and 4 with mixtures containing equal amounts of pigeon erythrocytes (Prbc) coupled to para-azobenzenearsonate (AA) and to para-azobenzene-N-trimethylammonium (TMA). On day 6, the allotypes of antibody from plaque-forming cells (PFC) of the blood were determined by observing the inhibition of plaque formation by anti-allotype sera. Anti-AA PFC appeared to consist for the most part of cells making antibody of allotype a1 since 65% of them were inhibited by anti-a1 serum and only 8% by anti-a3. Anti-TMA PFC, on the other hand, appeared to consist mostly of cells making antibody of allotype a3, since less then 1% of them were inhibited by anti-a1 but 47% by anti-a3. Antibody allotype for spleen PFC was also determined on day 6 and was similar to that found for blood PFC. Anti-AA PFC were inhibited 74% by anti-a1 serum and 15% by anti-a3 whereas anti-TMA PFC were inhibited 19% by anti-a1 and 43% by anti-a3.

Serum hemolysin specific for AA hapten from a1a3 animals was also strongly inhibited by anti-a1 serum but not by anti-a3 whereas the converse was true for hemolysin against TMA hapten.

The a1a3 rabbits, in which the anti-AA was restricted to allotype a1, were mated to produce homozygous a3a3 animals. When the PFC and serum antibodies of these a3a3 offspring were examined by specific inhibition, the anti-AA activity was found to be of allotype a3 rather than being anegative.

The number of anti-AA PFC in the blood of a3a3 rabbits was lower than that in blood of a1a3 or a1a1 animals. In addition, the TMA hapten appeared to inhibit the response to the AA hapten. Thus a1a3 rabbits immunized with AA-Prbc alone had 14-fold more anti-AA PFC or 18-fold higher anti-AA hemolysin titer than a3a3 animals immunized with both AA-Prbc and TMA-Prbc.

Our results are discussed in relation to various explanations which have been offered for an imbalance of allotypes in a given antibody.