bluuebunny/crossref_metadata_2025 huggingface dataset:
Clonal Nature of the Immune Response to Phosphocholine
Authors: Claflin J. Latham , Cubberley Marge
Published: December 2022

The molecular heterogeneity of IgG antibodies to phosphocholine (PC) having a defined idiotype was examined in BALB/c mice immunized with PC-keyhole limpet hemocyanin (KLH). Specific antibodies were separated by isoelectric focusing in polyacrylamide gels and characterized for PC-binding, idiotype, and isotype by direct in situ labeling with 125I-labeled reagents followed by autoradiography. After immunization with PC-KLH, BALB/c produce 20 to 100 µg/ml of IgM and 80 to 300 µg/ml of IgG anti-PC antibody. The dominant fraction of anti-PC antibodies in BALB/c (and a lesser fraction in other strains) possesses idiotypic determinants found on a PC-binding myeloma, TEPC15. Among 65 BALB/c examined, all produced an identical spectrotypic pattern of antibodies possessing T15 idiotypic determinants. Three major sets of T15-idiotype bearing bands were observed, but they belonged to three different IgG subclasses: IgG1, IgG2, and IgG3. These data support the germ line origin for this dominant set of antibodies in the anti-PC repertoire of BALB/c and indicate that they arise from a single rather than multiple VH-VL pairs.
The Induction of Cytolytic T Lymphocytes with Specificity for p-Azophenylarsonate Coupled Syngeneic Cells
Authors: Sherman Linda A. , Burakoff Steven J. , Benacerraf Baruj
Published: December 2022

Conditions are described for diazonium linkage of the hapten p-azophenylarsonate (Ar) to mouse lymphocytes for the purpose of induction of syngeneic cytolytic T lymphocytes. These cytolytic effector cells do not lyse target cells that are coupled with an unrelated hapten, trinitrophenyl. Cell lysis can be blocked by the addition of anti-Ar antiserum. Only those target cells that share H-2 K or D end antigens with the stimulating cell population are susceptible to lysis. Several methods were used in an attempt to demonstrate the presence of the Ar cross-reactive idiotype on CTL induced in A/J mice.
Isolation and Antigenic Characterization of the Product of a Third Polymorphic H-2 Locus, H-2L
Authors: Hansen Ted H. , Sachs David H.
Published: December 2022

The serology, immunochemistry, and genetics of the product(s) of a third H-2 locus, H-2L (previously designated D') have been studied by using an antiserum raised in BALB/c-H-2db mutant mice against tissues from the wild type strain, BALB/c. Genetic mapping studies and sequential immunoprecipitation experiments both indicate that this antiserum reacts specifically with L molecules. These results imply that an H-2L product is anti-genically undetectable in BALB/c-H-2db mice and that the lesion in this mutant is confined to the H-2L and not the H-2D locus. Two new specificities, H-2.64 and H-2.65, are defined by the reactivity of anti-L serum on allogeneic cells, and the strain distribution of these specificities suggests the existence of at least three H-2L alleles. This third H-2 locus is therefore polymorphic and in view of this and other similarities to the H-2K and H-2D loci, it must be considered in any evolutionary models dealing with the origin of multiple subloci of the major histocompatibility complex.
Effect of Ultraviolet Light on Dinitrochlorobenzenespecific Antigen-Presenting Function
Authors: Levis William R. , Lincoln Paula M. , Dattner Alan M.
Published: December 2022

HLA-A and -B antigens are resistant to ultraviolet (UV) light whereas HLA-D, as defined by the mixed leukocyte culture reaction (MLR), is sensitive to UV light in initial or first culture. In primed LD typing, the HLA-D responsible for stimulation is relatively resistant to UV light in second culture. We have developed a method for evaluating a specific proliferative response to DNP in dinitrochlorobenzene (DNCB)-sensitized human subjects and found a requirement for a component of autologous leukocytes to activate DNP-responsive lymphocyte clones. Autologous DNBSO3-coupled leukocytes with and without UV were used to stimulate lymphocytes in DNCB-sensitized subjects. 3HTdR incorporation was measured after 4 or 5 dys in 1st or “primary” culture. After 9 to 14 days, blast-derived lymphocytes were set up in 2nd or “secondary” cultures and exposed to the same autologous DNBSO3-treated leukocytes with and without UV. Accelerated 3HTdR incorporation was measured in these secondary clonally primed cultures after 20 to 44 hr. The results showed that UV light abrogated the antigen-presenting or stimulator function in the 1st culture and diminished, but did not abrogate, the antigen-presenter function in the 2nd culture. These findings indicate that the DNP-antigen presenting function is behaving in a manner similar to that observed for mixed leukocyte culture determinants in 1st and 2nd cultures.
Characteristics of Complement Receptor-Bearing Cells in the Spleens of Tumor-Bearing Mice
Authors: Epstein Robert S. , Lopez Diana M. , Sigel M. Michael , Ross Gordon D.
Published: December 2022

In the course of mammary tumor development, a population of nylon nonadherent cells with CR appears in the spleens of tumor-bearing mice although none are ever detected in normal mice. These cells apparently arise in response to immunologic stimulation. In a series of studies we have further characterized subsets of T cells (CR+ and CR-) with regard to their responses to mitogens in the lymphocyte transformation assay. Nylon column nonadherent cells from the spleens of tumorbearing mice were rosetted in a complement receptor assay using EAC rosetting, and CR+ cells were separated from CR- by centrifugation in a discontinuous Ficoll gradient. CR+ T cells responded strongly to PHA and Con A and in addition responded to LPS, an activity not usually associated with conventional T cells. In contrast, CR- T cells from tumor-burdened mice responded to PHA but failed to respond to Con A or LPS.
Fc-Receptors on Human T Lymphocytes
Authors: Pichler Werner J. , Lum Lawrence , Broder Samuel
Published: December 2022

T lymphocytes of 11 healthy humans were enriched and studied for Fc-IgG or Fc-IgM receptor expression by using a rosette technique, namely bovine red cells coated with rabbit IgG {EA-IgG} or IgM {EA-IgM}. Eighty percent (±9.2) of the T cells (mean ± S.D.) exhibited Fc-IgM receptors after culture (designated Tµ) and 22% (±9.4) expressed Fc-IgG receptors (designated Tγ), the latter occasionally decreasing after culture at 37°C. The two populations could be readily separated by interaction of Tγ cells with EA-IgG and Ficoll-Hypaque gradient separation. In the interface we found T non-γ cells, which consisted of over 90% Tµ cells and about 1% Tγ cells. In the pellet an enriched Tγ cell population was found, 79% (±8). After lysis of the bound EA-IgG, incubation of the cells at 37°C led to capping, endocytosis, and/or shedding of the Fc-IgG receptor. Culture of these cells led to expression of Fc-IgM receptors on these originally Tγ cells, whereas only a few cells re-expressed Fc-IgG receptors. The transition of Tγ → Tµ cells was also observed in T cells precultured for 24 hr and required immune complex (IC) interaction. Adding sodium azide or maintaining the cells at 4°C blocked Fc-IgM receptor expression. X-irradiation, cytochalasin B, and colchicine had no effect. Pronase treatment of positively selected Tγ cells uncovered Fc-IgG receptors, which did not cap, and after 40 hr in culture, T cells bearing Fc-IgG and Fc-IgM receptors were found. Therefore, Fc-IgG and Fc-IgM receptors do not appear to be markers for distinct T cell subsets under some conditions. Rather, a subset of T cells may conceivably express both classes of receptors at different functional stages. The interaction of Fc-IgG receptors with IC may serve to alter the overall expression of Fc-receptors.
Antibody-Independent Neutralization of Vesicular Stomatitis Virus by Human Complement
Authors: Mills Bonnie J. , Cooper Neil R.
Published: December 2022

Normal human serum neutralized up to 5 logs10/ml of vesicular stomatitis virus (VSV) grown in hamster (BHK), mouse (L), or human (HFF) cell lines when equal volumes of virus (∼105 PFU/ml) and serum were incubated together for 30 min at 37°C. The neutralizing activity of fresh serum was heat labile, thus distinguishing this activity from that of immune human serum which was heat stable. The lack of an antibody requirement for VSV neutralization by normal human serum was demonstrated in three ways: first, isolated human immunoglobulins failed to neutralize VSV or potentiate neutralization by fresh serum; second, absorption of fresh serum with uninfected cells or cells bearing VSV antigens did not reduce neutralizing activity; third, agammaglobulinemic serum mediated C3b deposition on the viral surface. Neutralization by normal human serum was complement (C) dependent and required the early reacting components of the classical C pathway (C1q, C2, C4) but not the late reacting components (C5, C6, C7, C8) or the alternative C pathway (factor B). Incubation of serum with concentrated VSV reduced the CH50, C2, C3, and C4 titers by 23.5, 21, 22, and 31%, respectively, and was accompanied by deposition of C3b on the viral surface. Purified VSV and the isolated external glycoprotein of VSV, G protein, bound 125I-C1q, and purified VSV also bound C1̄. However, VSV did not activate purified reconstituted C1, and VSV was not neutralized by isolated C1, C2, C3, C4, and C5. Therefore, an additional serum factor is apparently required for neutralization.

These studies indicate that neutralization of VSV by normal human serum involves the components of the classical C pathway through C3b and a nonimmunoglobulin serum factor. The reaction is initiated by the direct attachment of C1 to the G protein, which leads to C activation and the sequential deposition of the C components through C3b on the viral envelope. The mechanism of action of the nonimmunoglobulin serum factor remains to be established.
A Survey of Murine Ia Antigens Among Populations of Texas Wild Mice
Authors: Duncan William R. , Wakeland Edward K. , Klein Jan
Published: December 2022

The polymorphism of two serologically detected loci in the I region was assessed in wild mice trapped in six separate locations in Texas. About 100 mice were typed by the two-stage microcytotoxicity test with a battery of alloantisera detecting 10 private, semi-private, or public specificities in either the I-A or I-EC loci. This survey has revealed that three different private I-region specificities are present among wild mice in Texas at frequencies characteristic of stable polymorphisms. Two of these private specificities, Ia-1.2 and Ia-1.4, were also present in the descendents of mice trapped in Michigan, indicating that these alleles are widely distributed among wild mice in the United States. A comparison of the distribution of I region specificities with K or D region specificities indicated that linkage disequilibrium among these H-2 loci was a rare, but detectable event. Finally, breeding studies with wild, male mice indicate that about 90% of the wild males are heterozygous for H-2 antigens.
Expression of I-Subregion Encoded Antigens on Accessory Cell Populations
Authors: Ahmann G. B. , Singer A. , Dickler H. B. , Sachs D. H. , Hodes R. J.
Published: December 2022

We have investigated the expression of Ia antigens on accessory cells which function in two murine response systems: the proliferative response to Con A and the in vitro primary antibody response to soluble TNP-conjugated protein antigens. Pretreatment of murine lymphoid cells with anti-Ia and complement (C) abrogated their ability to respond to Con A. Reconstruction experiments have shown that glass adherent spleen cells treated with a T cell-specific rabbit anti-mouse brain serum and C and irradiated with 1000R (spleen-adherent cells, SAC) were able to restore responsiveness to Con A. In a second assay system, T cell-dependent IgM responses of normal spleen cells to soluble TNP-conjugated protein antigens were abrogated by Sephadex G-10 passage. Responses in this system were similarly reconstituted by the addition of non-T, non-B, radioresistant SAC as accessory cells.
Detection of Two Independent Ia-Like Antigens Determined by Genes Linked to Rat MHC
Authors: Shinohara Nobukata , Sachs David H.
Published: December 2022

The mouse alloantiserum B10.D2 anti-B10.BR (H-2d anti-H-2k) crossreacted with rat lymphocyte surface glycoproteins with characteristics of Ia antigens. Sequential precipitation analysis on solubilized radiolabeled Lewis rat spleen cell alloantigens pretreated with BN anti-Lewis (Ag-B3 anti-Ag-B1) alloantiserum revealed that the Ia-like antigens detected by the mouse alloantiserum also reacted with the rat anti-Ia antibodies. The rat alloantiserum also detected another set of Ia-like antigens which did not crossreact with the mouse alloantibody. Precipitation analysis using congenic rat strains confirmed that all Ia-like antigens precipitated by the rat alloantibody were encoded by Ag-B-linked genes. Thus the shared Ia-like antigen must also be the product of Ag-B-linked gene(s) or be physically associated with such products. The existence of two separable Ia antigens suggest the existence of more than one sublocus coding for Ia antigens within the rat MHC. The determinant detected on the crossreactive molecule by the mouse alloantibody is probably distinct from that detected by the rat alloantibody since the mouse alloantibody reacted similarly with both Lewis and BN lymphocytes.
Comparison of Primary Structures of Ia Products from the I-A and I-E/C Subregions
Authors: Freed John H.
Published: December 2022

The primary structures of α and β chains from both I-A and I-E/C subregions of several haplotypes were exchange chromatography of tryptic peptides. The materials examined were obtained from NP-40 extracts of biosynthetically radiolabeled normal spleen cells. The preparations were subjected to LcH-Sepharose affinity chromatography prior to immunoprecipitation with specific alloantisera; α and β chains were separated by polyacrylamide gel electrophoresis and/or hydroxylapatite chromatography in SDS. Comparative chromatographies of tryptic peptides from α and β chains reveal that corresponding chains from the highly cross-reactive I-Ak and I-Ar gene products share approximately 65–75% of their peptides. Preliminary results suggest that for the I-E/Ck and I-E/Cr products the α chains share all peptides, whereas the β chains share only 65% of their peptides. This latter result, considered in light of the proposed assignment of Ia.7 to the I-E subregion (David and Cullen, in press), suggests that the I-Ek specificity Ia.22 (which I-Er lacks) may be carried specifically on the Eβk chain.
Different Antibodies Made to Myoglobin Under Control of Distinct Ir Genes in Different I-Subregions of H-2
Authors: Berzofsky Jay A. , Killion Douglas J.
Published: December 2022

Two Ir genes in distinct I-subregions of H-2 have been shown to control the antibody response to sperm whale myoglobin (Mb) in mice (Berzofsky, J. A., J. Immunol. 120, 360, 1978). B10.D2 (H-2d) mice (high responders) have both Ir-Mb-1 (in I-A) and Ir-Mb-2 (probably in I-C), while B10 (H-2b) and B10.BR (H-2k) mice (low responders) have neither. Recombinant strains B10.A and B10.A(5R) with only Ir-Mb-2 are intermediate in response to whole Mb. However, when the same sera were tested for antibodies binding to fragment (132–153), B10.A 3° sera, which had 6-fold more anti-Mb than B10.BR sera, had no more antibody to this fragment (whereas the B10.D2 sera had 10-fold more anti-(132-153)). Similarly, the B10.A(5R) sera had barely more anti-(132-153) than B10 sera, even though they had 4-fold more total anti-Mb. Conversely, D2.GD mice (with only Ir-Mb-1) made as much anti-(132–153) in the anti-Mb sera as did congenic DBA/2 mice, with both genes.
Genetic Control of the T-Lymphocyte Proliferative Response to Cytochrome c
Authors: Solinger A. M. , Schwartz R. H. , Ultee M. , Margoliash E.
Published: December 2022

Cytochromes c have been used as antigens in a murine T-lymphocyte proliferation assay in order to characterize the nature of determinants whose recognition is under immune response (Ir) gene control. In the present studies, the T-cell proliferative response to pigeon cytochrome c was shown to be under the control of two complementing MHC-linked Ir genes in mice of the H-2a and H-2k haplotypes, while eight other haplotypes tested were low or non-responders. Complementation was demonstrated by showing that an F1 hybrid between two non-responder recombinant strains (H-2h4 and H-2i5) could respond to pigeon cytochrome c. The determinant on the cytochrome recognized in this immune response was located to the C-terminal portion of the molecule around residue 100. This was shown by the failure or success of closely related cytochromes to cross-stimulate T-cells immune to pigeon cytochrome.
Evidence for Ir Gene Expression in the Antigen-Presenting Cell
Authors: Yano Akihiko , Schwartz Ronald H. , Paul William E.
Published: December 2022

Proliferative responses by peritoneal exudate T-lymphocyteenriched cells (PETLES) from GAT primed mice can be stimulated by GAT-pulsed spleen cells from non-immune animals only if the donors of both cell types are identical at the I-A subregion of the major histocompatibility complex (MHC). However, in 4 separate (high responder × low responder)F1 strains, GAT-primed PETLES could be stimulated by GAT/presenting cells from either the F1 or the high responder parent but not by cells from the low responder parent. Addition of equal numbers of GAT-pulsed low responder spleen cells did not affect the response of F1 PETLES to GAT on high responder spleen cells. Furthermore, pre-treatment of F1 PETLES with anti-Ly 2.2 serum and complement did not allow these cells to respond to GAT-pulsed low responder parental spleen cells. This suggests that the failure of spleen cells from low responder parental strains to present GAT to F1 PETLES is not due to the presence of suppressor cells in either the antigen-presenting or the responding cell populations.
H-2, Thymus and H-2 Restricted T Cells
Authors: Zinkernagel Rolf M.
Published: December 2022

The radioresistant portion of the thymus dictates the H-2 self specificity spectrum of T cells and the peripheral lymphoreticular cells' H-2 type determines which of the thymically selected specificities for H-2 are to be expressed by antigen specific effector T cells; these contentions derive from experiments with irradiation bone marrow and thymus chimeras. These phenomena cannot be readily explained by suppression as shown in mixture experiments. Furthermore, specificity for thymic self-H-2 is exquisite, since lymphocytes from C57BL/6 (Kb) → Hz170 (Kba) mutant mice chimeras do not cross-react on Kb. These findings have great theoretical and practical implications with respect to reconstitution of immunocompetence, Ir-gene phenomena and our understanding of T cell recognition.
Natural Cytotoxic Cells against Solid Tumors in Mice
Authors: Stutman Osias , Paige Christopher J. , Figarella Elizabeth Feo
Published: December 2022

Lymphohemopoietic cells from normal mice show cell-mediated cytotoxicity (CMC) against syngeneic or allogeneic chemically induced fibrosarcomas, when tested in a 24-hr cytotoxicity assay with 3H-proline pre-labeled adherent target cells. When two BALB/c tumors, Meth A, and Meth 113 were studied, spleen cells from syngeneic normal donors showed high CMC against Meth A and no CMC toward Meth 113. However, Meth 113 was susceptible to natural CMC by spleen cells from other mouse strains or PHA-induced CMC by syngeneic spleen cells. Other solid tumors tested showed variable patterns of susceptibility-resistance to natural CMC, whereas normal fibroblasts were negative. Susceptibility to natural CMC does not correlate with expression of murine leukemia virus-related antigens. Established lines are more susceptible to natural CMC than secondary tumor cultures, although susceptibility-resistance appear as stable properties of the targets. All the mouse strains and sublines tested showed natural CMC against the Meth A target; however, only a few strains (A/J, CBA/HN, NZB, and C3H/He) showed natural CMC against Meth 113. Natural CMC against Meth 113 of (BALB/c × A/J) F1 spleen cells is dominant, and the hybrid behaves like the reactive A/J parent. Natural cytotoxic (NC) cells are detected in nude mice in a BALB/c or CBA/H background (and also in CBA/HN mice, which have a B cell defect). The tissue distribution of NC cells shows high activity in spleen and marrow, with lower activity in lymph nodes, peritoneal washings, and thymus. Peritoneal exudates induced by complete Freund's adjuvant or BCG show high NC cell activity. NC activity in spleen was detected from birth and does not seem to decay with age (even at 1 or 2 years of age). The described characteristics of the NC cell have some similarities (and some discrepancies) with the natural killer (NK) cell described against T lymphoma targets. The differences include: 1) strain distribution; 2) appearance at birth; 3) no decay with age, and 4) to some extent, tissue distribution (i.e. both systems show high activity in spleen; however, NC cells are detected in thymus whereas NK cells are not). The similarities include: 1) susceptibility not related to expression of murine leukemia virus antigens; 2) no H2 restriction of cytotoxicity; 3) activity present in nude mice (and CBA/HN); 4) reactivity dominant in F1 hybrids, and 5) increase activity in peritoneal exudates after BCG administration. It seems possible that these systems may act as anti-tumor surveillance devices.
Tumor-Associated Antigens of Rat Moloney Sarcoma Cells
Authors: Leung Kiu , Jones Joe M. , Feldman Joseph D.
Published: December 2022

The dissociated cell surface membranes of a rat Moloney sarcoma (MST), derived from a BN rat, were extracted with 2 M KI, with 6 M guanidine thiocyanate, or by papain digestion. Extracts obtained with these three reagents were fractionated on columns of controlled-pore glass, 170 Å pore size. A fraction was eluted from each preparation that contained tumor-associated antigens (TAA), viral, fetal, and histocompatibility antigens. With an antibody specific for TAA, the TAA, devoid of detectable viral, fetal, and histocompatibility antigens, were co-precipitated by addition of goat antibody to rat immunoglobulin. After electrophoresis, on slab gels, three bands were detected with estimated m.w. of 185,000, 150,000, and 70,000. No such bands were detected on slab gel electrophoresis with extracts of BC5, a chemically induced tumor, of normal BN lymphoid cells, of M-MuLV, or of fetuses, after incubation with anti-TAA antibody and goat antibody to rat immunoglobulin. TAA extracts prepared with 2 M KI, 6 M guanidine thiocyanate, or papain digestion showed immunologic reactivity. Cold TAA inhibited the co-precipitation of labeled TAA by rat antibody specific for TAA; they elicited antibody in guinea pigs but not in rats; and antibodies specific for TAA were cytotoxic to MST in the presence of C.
Micro-Leukocyte Adherence Inhibition
Authors: Russo Anthony J. , Howell John H. , Leveson Stephen H. , Douglas Holyoke E. , Goldrosen Martin H.
Published: December 2022

To study the cellular basis for specific antigen-induced leukocyte adherence inhibition, enriched populations of B cells, T cells, and monocytes were prepared by a two-stage adherence separation procedure from spleen cells of normal C57BL/6J mice and mice bearing progressively growing MCA-38 tumors. The reactor cell undergoing specific antigen-induced adherence inhibition was identified as a monocyte (esterase positive, did not respond to mitogens, and did not bear Thy 1.2 antigen or surface immunoglobulin). Furthermore, an enriched population of MCA-38 sensitized B cells could program normal monocytes to undergo specific antigen-induced adherence inhibition. This programming could be abolished by pretreatment of the MCA-38 sensitized B cells with anti-immunoglobulin and complement (indirect cytotoxicity method). In contrast, enriched populations of MCA-38 sensitized T cells could not program normal nylon wool adherent cells to undergo antigen-specific adherence inhibition; and anti-Thy 1.2 serum and complement had no effect on specific antigen-induced adherence inhibition. Thus, in this murine tumor model, leukocyte adherence inhibition appears to be due to the programming of monocytes by sensitized B cells.
Specific Inhibition of Cytotoxic Memory Cells Produced against UV-Induced Tumors in UV-Irradiated Mice
Authors: Thorn Richard M.
Published: December 2022

Cytotoxic responses of UV-irradiated mice against syngeneic UV-induced tumors were measured by using a 51Cr-release assay to determine if UV treatment induced a specific reduction of cytotoxic activity. The in vivo and in vitro primary responses against syngeneic tumors and allogeneic cells were unaffected, as was the “memory” response (in vivo stimulation, in vitro restimulation) against alloantigens. In contrast, the memory response of UV-treated mice against syngeneic, UV-induced tumors was consistently and significantly depressed. The cytotoxicity generated by tumor cells stimulation in vivo or in vitro was tumor-specific and T cell-dependent. Since the primary response against syngeneic UV-induced tumors produces apparently normal amounts of tumor-specific cytotoxic activity, UV-treated mice may not reject transplanted syngeneic tumors because of too few T effector memory cells. These results imply that, at least in this system, tumor rejection depends mostly on the secondary responses against tumor antigens and that at least one carcinogen can, indirectly, specifically regulate immune responses.
Immunologic Functions of Ia-Bearing Epidermal Langerhans Cells
Authors: Stingl Georg , Katz Stephen I. , Clement Loran , Green Ira , Shevach Ethan M.
Published: December 2022

Langerhans cells (LC) constitute a morphologically well-characterized minor subpopulation of the mammalian epidermis whose functional role is still a matter of conjecture. The hypothesis that LC represent an epidermal equivalent to cells of the monocyte-macrophage-histiocyte series is supported by the recent observations that in humans and guinea pigs LC are the only epidermal cells that express Fc-IgG receptors, C3 receptors, and Ia antigens. Using inbred strain 2 and strain 13 guinea pigs, we investigated in this study whether LC can mediate the same immunologic functions as Ia-bearing macrophages. LC-enriched and LC-depleted epidermal cells were prepared by separation of Fc-IgG rosetting epidermal cells on density gradients. When both populations were tested for the biosynthesis of alloantigens by immunoprecipitation techniques, Ia antigen synthesis was restricted to the LC-enriched fraction. Functional studies demonstrated that antigen-pulsed LC-enriched epidermal cells induce a proliferative response in immune T cells that is comparable in magnitude to that seen with macrophages. Moreover, effective presentation of immunologically relevant antigen requires syngeneity between LC-enriched epidermal cells and responder lymphocytes. In the mixed leukocyte reaction (MLR), LC-enriched epidermal cells were as effective stimulators as macrophages. LC-depleted epidermal cells, by contrast, induced little or no stimulation in both assay systems. Both the antigen-presenting and the MLR-stimulatory capacities of LC-enriched epidermal cells could be abrogated by pretreatment with anti-Ia sera and complement. The presence in the epidermis of Ia-bearing LC, capable of mediating the immunologic functions of Ia-bearing macrophages, has important clinical implications with regard to the role of LC as sensitizing cells in both contact hypersensitivity and skin graft rejection.
Differential Effect of Neonatal Injections of Anti-µ or Anti-δ Antibodies on the Synthesis of IgM, IgD, IgE, IgA, IgG1, IgG2a, IgG2b, and IgG2c Immunoglobulin Classes
Authors: Bazin Herve , Platteau Bernadette , Beckers Andree , Pauwels Romain
Published: December 2022

The effect of neonatally initiated injections of anti-µ or anti-δ antibodies on IgM, IgD, IgA, IgE, IgG1, IgG2a, IgG2b, and IgG2c immunoglobulin subclasses are described. As in chickens and mice, anti-µ suppression was demonstrated to be effective on rat IgM, IgA, and IgG classes but also on IgD and IgE classes and on the four subclasses of IgG. The anti-δ suppression clearly affected the IgD synthesis but also that of IgE. The different IgG subclasses were more or less diminished, but, on the contrary, the IgG2c serum level from day 55 on was largely increased. The IgM and IgA synthesis both were slightly affected with respect to their serum levels but significantly at the membrane level.

Rats suppressed by anti-µ or anti-δ sera were unable to build up a reaginic immune response taken as an example of T-dependent immune response.

The anti-µ suppression on all the immunoglobulin classes confirms that the IgM receptors are the first to appear on the precursors of the B lymphoid cells. The IgD suppression also confirms that the majority of B lymphoid cells carry IgD receptors during their maturation. Moreover, the absence of reaginic immune response indicates that the lack of the IgD receptor can make animals unable to produce primary IgE immune response.

The IgD receptor may be considered as an unspecific isotype-positive signal necessary for the majority of the B lymphoid cells to react for the first time to a specific antigen stimulation. The differential results of the anti-δ suppression, contrary to the general suppression obtained by the anti-µ inoculation, could be explained by the more or less important requirement by each isotype cell line, to be stimulated primarily by an antigen with the help of IgD class receptors.
An Attempt to Raise Antibodies against 2-O-α-D-Galactopyranose, the Disaccharide Unit of Collagen
Authors: Ebisu Shigeyuki , Garegg Per J. , Iversen Tommy , Goldstein Irwin J.
Published: December 2022

The p-isothiocyanatophenyl β-glycoside of 2-O-α-D-glucopyranosyl-D-galactopyranose, the disaccharide unit of collagen, was coupled to bovine serum albumin for use as an artificial antigen. The conjugate precipitated with concanavalin A but not with the Ricinus communis lectin. Rabbit antisera raised against the conjugate were characterized by immunodiffusion, quantitative precipitation, and hapten inhibition. After absorption with bovine serum albumin the antisera exhibited a specificity for the introduced p-isothiocyanatophenyl 2-O-α-D-glucopyranosyl-β-D-galactopyranoside. The relative importance of an α-configuration and the ordered sequence of the two sugars for binding to antibody were observed. The antibody-combining site was shown to involve the aglycone portion of the introduced hapten. 5-O-(2-O-α-D-glucopyranosyl-O-β-D-galactopyranosyl)-hydroxylysine and 5-O-(2-O-α-D-glucopyranosyl-O-β-D-galactopyranosyl)-hydroxylysine peptide, the glycopeptide unit in collagens, were poor inhibitors of the conjugate-antibody precipitation system. However, acetylation of the ε-amino groups of the hydroxylysyl residues converted these noninhibitory substrates into considerable inhibitors, suggesting that a charged -NH3+ group may interfere with binding of the haptenic groups to antibody.
Modulation of the Antigenicity of C1r̅ and C1s̅ by C1̄ Inactivator
Authors: Ziccardi Robert J. , Cooper Neil R.
Published: December 2022

The first complement component, C1, was activated in normal human sera by addition of aggregated IgG, immune complexes, and lipopolysaccharide, after which the concentrations of the C1 subcomponents, C1r and C1s, were measured by single radial immunodiffusion. A progressive reduction in C1r antigen content was observed as greater amounts of activator were added to normal human sera. Three lines of evidence proved that the apparent disappearance of C1r was due to interaction of C1r̅ with C1̄ Inactivator (C1̄ In). First, no loss of C1r antigen was detected in sera from patients with hereditary angioneurotic edema, which lack functional C1̄ Inactivator. Second, reconstitution of these patients' sera with physiologic concentrations of purified C1̄ In resulted in the disappearance of C1r antigen. Third, purified C1̄ In greatly reduced the intensity of the precipitin line formed between purified C1r̅ and anti-C1r̅.

In parallel experiments, certain antisera to C1s were found to show a comparable disappearance of C1s antigenic content in activated human sera. This antigenic loss was similarly shown to be due to the interaction of C1s̅ with C1̄ In. However, it should be noted that the detection of changes of C1s̅ antigenicity was dependent on the specificity of the C1s antisera employed, since antisera with antigenic specificities apparently directed to determinants on the C1s molecule distant from the region of interaction with C1̄ In did not show changes with activation.

Ultracentrifugal studies demonstrated that C1r̅ and C1s̅ formed soluble complexes with C1̄ In in activated serum, thus precluding the possibility that C1r̅ and C1s̅ escape immunochemical detection in activated serum by being present in large aggregated complexes. Also ruled out, in the case of C1s̅, is the possibility that the binding of C1̄ In leads to a major conformational rearrangement of C1s̅ with resulting changes in exposed antigens of the molecule. It is most likely that the loss in C1r̅ and C1s̅ antigenic content with activation of C1 in serum occurs because of simple physical masking of the relevant antigens of C1r̅ and C1s̅ by the relatively large C1̄ In molecule.
β-Estradiol Reduces Natural Killer Cells in Mice
Authors: Seaman William E. , Blackman Marcia A. , Gindhart Thomas D. , Roubinian Jirayr R. , Loeb John M. , Talal Norman
Published: December 2022

β-estradiol was administered to mice continuously by diffusion from a silastic tube that was implanted subcutaneously at 4 weeks of age. Four to 6 weeks of estrogen administration caused a substantial reduction in natural killer cell activity in the spleens from mice of either sex. Androgen (5α-dihydrotestosterone) did not. Castration of male or female mice did not affect natural killing and did not alter the effect of β-estradiol. Estradiol did not affect natural killing in vitro and the loss of natural killing was not due to a soluble or a cellular suppressor of natural killing. The effects of estradiol were not dependent on the thymus, since estradiol reduced natural killing in mice that had been neonatally thymectomized. After removal of the estrogen implant, natural killing recovered over a period of 8 weeks. The loss of natural killing may reflect a loss of bone marrow secondary to estrogen-induced osteosclerosis.
Autoantibody to a Nuclear Antigen in Proliferating Cells
Authors: Miyachi Kiyomitsu , Fritzler Marvin J. , Tan Eng M.
Published: December 2022

This study reports the identification of an autoantibody in the sera of some patients with systemic lupus erythematosus that reacted with nuclear antigen(s) of proliferating cells. The autoantibody was initially detected by the observation that it did not react in immunofluorescence with nuclei of renal tubular or glomerular cells, nor with hepatic parenchymal cells, but only reacted with scattered cells in the interstitial tissue of these two organs. In contrast, many lymphocytes in lymph node follicles, spleen, and thymus reacted with this antinuclear antibody. Normal peripheral blood lymphocytes did not show nuclear staining but after mitogenic stimulation, 20% of cells became positive. Nuclear staining was not restricted to lymphocytes but was also observed in several tissue culture cells lines such as Hep-2 cells (human epithelial carcinoma), Ehrlich ascites tumor cells, and baby hamster kidney cells. The reactive nuclear antigen(s) was soluble in physiologic saline and reacted with serum autoantibody to give a precipitin line in immunodiffusion that was immunologically distinct from DNA and other known nuclear antigen-antibody precipitating systems. Autoantibodies to proliferating cell nuclear antigen(s) might serve as useful biologic markers to study stimulated lymphocytes and other proliferating cells.
Macrophage-T Cell Interactions in the Con A Induction of Human Suppressive T Cells
Authors: Raff Howard V. , Cochrum Kent C. , Stobo John D.
Published: December 2022

Macrophage-T interactions are required for the Con A-induced generation of human Ts capable of inhibiting PHA-induced blastogenesis among autologous PBMC. Con A treatment of adherent cell-depleted PBMC, or PBMC recovered after a 7-day incubation in FCS, failed to generate Ts. Addition of adherent cells to either of these populations restored Con A inducible Ts. Discontinuous density gradient fractionation of adherent cells demonstrated that the required accessory cell was a low density macrophage bearing the human equivalent of murine Ia.
A New B Cell Differentiation Antigen (BDA) on Normal and Leukemic Human B Lymphocytes That Is Distinct from Known DR (Ia-Like) Antigens
Authors: Balch Charles M. , Dougherty Patricia A. , Vogler Larry B. , Ades Edwin W. , Ferrone Soldano
Published: December 2022

A newly defined human B cell differentiation antigen, designated as BDA, has been defined and partially characterized. BDA is expressed on normal human B cells and lymphocytic leukemia cells at all stages of known differentiation (pre-B cells to plasma cells). It is distinct from DR (Ia-like) determinants and other known B cell surface constituents.
Genetic Identification of a Locus Linked to the Rat MHC That Codes for a Membrane Antigen Detectable with Cytotoxic Lymphocytes
Authors: Lynch David H. , Dewitt Charles W.
Published: December 2022

Cell-mediated cytolytic (CMC) responses resulted from immunizations between rat strains purported to be identical at Ag-B, the major histocompatibility complex (MHC), but differing at other loci not linked to Ag-B. In vivo-priming followed by secondary in vitro stimulation was required to generate a measurable CMC response as determined by a 51Cr-release assay. Neither in vivo nor in vitro stimulation alone was adequate. The CMC responses generated in a strain combination considered Ag-B identical (LEW·B3:BN) were specific for a determinant controlled by a gene linked to Ag-B, which has been designated Ag-L. The CMC response appears not to be restricted to syngeneity at Ag-B. In addition, the data presented demonstrate a recombinant between Ag-B and Ag-L, and suggest that the gene has failed to transfer with the MHC during the isolation of the LEW·B3 and F-344·B3 congenic strains.
Specificity Studies on the Proliferative Response of Thymus-Derived Lymphocytes to Influenza Viruses
Authors: Butchko Gregory M. , Armstrong Robert B. , Ennis Francis A.
Published: December 2022

Studies were undertaken to characterize the lympho-proliferative response to primary influenza virus infection and to determine the specificity of antigenic recognition by proliferative T cells obtained from mice infected with influenza virus. A proliferative response to A/Port Chalmers/1/73 virus (H3N2) was first detectable in spleen cells 5 days postinfection. Activity persisted in immunized mice for at least 12 months. The duration of immune responsiveness detectable in the 3H-thymidine incorporation assay contrasts with previously reported data indicating a relatively transient appearance of splenic T cell immunity detectable by 51Cr release cytotoxicity assays on influenza-infected target cells. These experiments clearly indicate that splenic T cells from mice infected with influenza virus proliferate in vitro when exposed to antigens of the infecting virus and that they can distinguish between A and B type influenza viruses. The proliferative T cell recognition of influenza viruses within the A type, however, appears to have a broad cross-reactivity, in contrast to the specificity previously shown by cytotoxic T cells. Data obtained from A/Port Chalmers/1/73 virus treated with various anti-influenza sera suggest that the hemagglutinin glycoprotein is of considerable importance in the antigenic recognition by, and stimulation of, influenza-immune proliferative T cells. These assays of the T cell responses to influenza infection should be of value in investigating the complex immune response of the host to influenza viruses.
Enterically Induced Immunologic Tolerance
Authors: Richman Lee K. , Chiller Jacques M. , Brown William R. , Hanson Donald G. , Vaz Nelson M.
Published: December 2022

Specific immune unresponsiveness was induced in inbred mice (BDF1) by the administration of soluble ovalbumin (OVA) by gastric intubation. Anti-hapten (DNP) responses likewise were specifically diminished when animals were fed autologous carrier (OVA or keyhole limpet hemocyanin). Adoptive transfer of spleen cells demonstrated that the tolerant state could be maintained in irradiated recipient mice, and specific anergy could be transferred to normal recipient animals. Adoptive suppression was mediated by T lymphocytes, as demonstrated by nylon wool fractionation and susceptibility of the cells to anti-Thy 1.2 and complement. Transferred B cells had neither suppressive nor augmentative effects. Enteric administration of OVA also specifically diminished antigen-induced DNA synthesis of primed lymph node T cells, although suppressor cells were not identified in the lymph nodes per se.
IgA-Fc Receptors on Mouse Lymphoid Cells
Authors: Strober Warren , Hague Norma E. , Lum Lawrence G. , Henkart Pierre A.
Published: December 2022

Receptors specific for IgA were sought on mouse lymphoid cells by using a rosetting technique in which cells are exposed to modified ox red blood cells (ORBC) coated with IgA myeloma proteins having specificity for the modified ORBC. It was found that 4 to 6% of unseparated spleen lymphoid cells (pre-incubated at 37°C overnight) formed rosettes with MOPC 315-coated TNP-ORBC, with QUPC 52-coated dextran-ORBC, and with SAPC 10-coated arabinogalactan ORBC, whereas modified ORBC alone or modified ORBC mixed with irrelevant myelomas did not form appreciable numbers of rosettes; the cells forming rosettes were not neutrophils as shown by appropriate staining techniques. Thirteen to 22% of spleen cells also formed rosettes with IgG-coated ORBC (either IgG rabbit anti-TNP-coated TNP-ORBC or rabbit IgG anti-ORBC-coated ORBC); however, by using similar techniques, no cells forming rosettes with IgM-coated ORBC could be found. The specificity of the IgA-binding sites thus demonstrated was shown in inhibition studies: IgA antigen-antibody complexes formed near equivalence or, indeed, purified IgA were found to inhibit rosette formation with IgA-coated ORBC, whereas IgG complexes or purified IgG were found to cause no such inhibition. These inhibition studies were buttressed by depletion studies in which removal of cells binding IgG-coated ORBC with rosetting procedures was shown to have little or no effect on the percentage of cells binding IgA-coated ORBC in the residual cell population. The cells capable of binding IgA-coated ORBC are at least in part T cells in that the binding of IgA-coated ORBC is a property of nylon-column passed cells containing <3% sIg positive cells; in addition, 2 to 3% of thymocytes bind IgA-coated ORBC. Finally, the IgA binding sites appear to be Fc-specific in that F(ab′)2 fragments of IgA do not block IgA-rosette formation. In summary, we have identified an IgA-Fc-specific binding site on mouse lymphoid cells, including mouse T cells.
Target-Effector Interaction in the Natural Killer (NK) Cell System
Authors: Roder John C. , Kiessling Rolf , Biberfeld Peter , Andersson Birger
Published: December 2022

We have previously shown that a subpopulation of nylon wool nonadherent, lymphoid cells in normal mice binds selectively to tumor cell targets that are sensitive to killing by natural killer (NK) cells. In the present report these target-binding cells (TBC) were enriched or depleted by velocity sedimentation and the isolation of single target-effector conjugates indicated that the majority of TBC killed their targets. Furthermore, the frequency of TBC correlated well (r = 0.86) with lysis in a population during kinetics experiments. TBC resembled small, resting lymphocytes with membrane specializations in the area of target cell contact as indicated by electron microscopy and cytochemical techniques. Target cell binding occurred before lysis in kinetics studies and regenerated in parallel with the lytic potential of a trypsinized population devoid of surface Ig or θ-bearing cells. Cell contact was prevented in the presence of EDTA whereas metabolic inhibitors or inhibitors of serine proteases suppressed lysis with no effect on the frequency of TBC. Conversely, an interferon-inducing agent elevated NK-mediated cytolysis with no effect on the level of TBC. These results suggest that i) the majority of TBC that we detect represents the NK cell and ii) lysis and binding are independently controlled events. A tentative model of NK-mediated lysis is proposed.
Rat Mast Cell-Eosinophil Interaction in Antibody-Dependent Eosinophil Cytotoxicity to Schistosoma Mansoni Schistosomula
Authors: Capron Monique , Rousseaux Jean , Mazingue Christine , Bazin Hervé , Carpon André
Published: December 2022

The involvement of IgG2a antibodies and mast cells in antibody-dependent eosinophil cytotoxicity suggested a possible interaction between mast cells and eosinophils for in vitro killing of Schistosoma mansoni schistosomula. Cell purification procedures showed that a minimum ratio of mast cells was required to obtain eosinophil cytotoxicity. The incubation of mast cells with heat-aggregated IgG2a immunoglobulins before addition to a mast cell-depleted eosinophil population induced a significant degree of inhibition of cytotoxicity. Similarly, the heat-aggregated IgG2a Fc fragment had a strong inhibitory effect whereas incubation of mast cells with Fab fragment failed to inhibit the cytotoxic effect. The Fc portion of IgG2a immunoglobulins therefore seemed to be involved in binding to the mast cell surface. Furthermore, it was demonstrated that soluble mediators released after mast cell activation either by compound 48/80, or by IgE, or IgG2a-dependent reaction had the same effect as intact mast cells. These observations suggest that the eosinophil-dependent cytotoxicity mechanism requires a signal provided by soluble mast cell mediators in addition to antibody.
C3-Mediated Cytoadherence
Authors: Landen Beate , Strunkheide Klaus , Dierich Manfred P.
Published: December 2022

Particles carrying C3 in a random distribution (Etan-C3) bound to C3 receptor (Raji+) cells independent of temperature and irrespective of whether the Raji cells were fixed with glutardialdehyde. In contrast, the reactivity of EAC43b, having grouped C3b in clusters, was dependent on temperature. The interaction with Raji cells was inhibited if the latter were treated with a fixative. The reaction of both Etan-C3 and EAC43b was a function of the concentration of C3 and C3b, respectively. The conclusion is drawn that for the interaction between C3/C3b-carrying particles and C receptor cells the receptors and the ligands have to be grouped in a similar typographic distribution, irrespective if this arrangement is provided already from the beginning or if it is obtained only by lateral movement of one or both reaction partners.