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Lymphoid function in F1 leads to parent chimeras: lack of evidence for adaptive differentiation of B cells or antigen-presenting cells.

Information was sought on whether B cells undergo abnormal differentiation in F1 leads to parent chimeras (irradiated parental-strain mice reconstituted with F1-hybrid bone marrow cells). As assessed by collaborative responses to sheep erythrocytes in vivo, three different types of T cells restricted to interaction with strain a H-2 determinants were shown to collaborate as effectively with heterologous F1 leads to b chimera B cells as with homologous F1 leads to a chimera B cells. This applied to both primed and unprimed B cells, to IgM- and IgG-antibody formation and to production of Ig allotype. Thus, unlike T cells, B cells from F1 leads to parent chimeras behaved indistinguishably from normal F1 B cells. F1 leads to parent chimeras were also examined for their capacity to present antigen to normal F1 T cells in vivo. The results suggested that the antigen-presenting cells in these chimeras were no different than in normal F1 mice. Collectively these data imply that, at least in the situation studied, raising F1 stem cells in a parental-strain environment has a marked effect on T-cell specificity but does not discernably influence the differentiation of B cells or macrophage-like cells.

Adaptation, Physiological

Antigen presentation in the murine T lymphocyte proliferative response. II. Ir-GAT-controlled T lymphocyte responses require antigen-presenting cells from a high responder donor.

The activation of T lymphocytes from poly (Glu60Ala30Tyr10)n (GAT)-primed donors by GAT-pulsed nonimmune spleen cells was shown to require identity at the I-1 subregion of the major histocompatibility complex. However, GAT-primed T lymphocytes from (responder x nonresponder) F1 hybrids could only be stimulated to proliferate by GAT bound to high responder or F1 spleen cells but not by GAT bound to spleen cells from the low responder parent. The failure of spleen cells from low responder parental strains to present GAT was shown not to be due to the presence of suppressor cells in either the antigen-presenting or the responding cell populations. These results indicate that control of antigen-presenting cell-T lymphocyte interactions is one site of Ir gene expression.

Animals

Antigen presentation in the murine T-lymphocyte proliferative response. I. Requirement for genetic identity at the major histocompatibility complex.

A method is described for stimulating proliferation in primed populations of murine T lymphocytes using antigen bound to mitomycin-C-treated spleen cells. This form of antigen presentation appears to be an active process because heat-killed spleen cells are ineffective, and because genetic similarity at the major histocompatibility complex (MHC) between the responder T cells and the presenting spleen cells is required for effective interactions. At all times examined, from day 3 to day 6 of the proliferative response, syngeneic spleen cells presented antigen better to peritoneal exudate T-lymphocyte-enriched cells (PETLES) than semisyngeneic F(1) spleen cells, which in turn could present antigen better than totally allogeneic spleen cells. Spleen cell mixing experiments demonstrated that these genetic restrictions were not the result of suppression by the ongoing mixed lymphocyte reactions (MLR) in the allogeneic and F(1) cases. Furthermore, incompatibility at the Mls locus generated a strong MLR but failed to prevent antigen presentation if the spleen cells and PETLES were compatible. Genetic mapping studies demonstrated that compatibility at only the I-A subregion of the MHC was sufficient for effective presentation of the antigen, dinitrophenylated ovalbumin. Compatibility at only the K region, or the K and D regions was not sufficient. These results support the concept that functional activation of primed, proliferating T lymphocytes requires the participation of gene products coded for by the I region of the MHC. This conclusion is consistent with a growing body of evidence which suggests that most T cells recognize antigen in association with MHC gene products.

Animals

Cancer Immune Responsiveness and MHC Class I Antigen Presentation: Mechanisms of Immune Escape and Immunotherapy Resistance in Gastrointestinal Cancers.

The Antigen Processing and Presentation Machinery (APM) is essential for immune surveillance by enabling the presentation of antigenic peptides to T lymphocytes and facilitating the elimination of infected or transformed cells. In cancer, the integrity of this process influences cancer immune responsiveness (CIR), defined as a tumour's capacity to be recognised by the immune system and respond to immunotherapy. Tumours with intact antigen presentation pathways are more likely to generate effective antitumour responses, whereas APM defects promote immune escape and therapeutic resistance. Cancer cells frequently evade immune detection through altered antigen processing or reduced expression of major histocompatibility complex (MHC) class I molecules, limiting tumour antigen presentation to cytotoxic T lymphocytes. These alterations are increasingly recognised as determinants of response to immune checkpoint inhibitors and potential predictive biomarkers. APM defects may be reversible or irreversible. Interferon-mediated signalling can restore MHC class I expression and T-cell cytotoxicity in some tumours, whereas permanent genomic alterations affecting human leukocyte antigen (HLA) class I genes, β2-microglobulin (β2-m), or interferon-γ (IFN-γ) pathway components can severely impair antigen presentation. Emerging evidence highlights four mechanistic levels of APM perturbation: peptide generation, peptide loading, MHC class I integrity, and epigenetic regulation. Each contributes to distinct patterns of immune evasion. This review examines how MHC class I alterations influence CIR and contribute to immune evasion and immunotherapy resistance in gastrointestinal malignancies, while discussing therapeutic strategies to restore or bypass APM deficiencies.

Humans

Soluble HLA antigens present in normal human serum.

HLA antigens have been detected in normal human serum by inhibition of monospecific HLA antisera in the complement-dependent cytotoxicity test. The antigens present in serum were the same as those found on the donor's lymphocytes. HLA-A9 was unique in that it was present in serum at inhibitory titers of 1:8 to 1:32 whereas the majority of A-and B-locus antigens were present at titers of 1:4 or less. Serum HLA-A9 antigens were found in the high density lipoprotein fraction with a molecular weight greater than 200,000. In the presence of detergent they have an apparent molecular weight of 86,000 which is reduced to 46,000 when treated with papain. These values are similar to those obtained for detergent-solubilized cellular HLA antigens.

Chromatography, Gel

Non-H-2 linked control of low versus high responses of antigen-induced lymph node cell proliferation: possible role for antigen-presenting cells.

Quantitative differences in the magnitude of antigen-induced proliferative responses of sensitized lymph node cells between low (C3H/Anf or C3H/Cr) and high (C3H/Hej or CBA/j) responder H-2k mice have been observed 1 to 2 weeks after in vivo sensitization with antigen (OVA, PPD, and GAT). We have shown that antigen presentation is less effective in the sensitized lymph node cell populations from low responder mice compared with those from high responder mice, suggesting that the number and/or functional status of antigen-presenting cells in the regional lymph nodes may be a key factor in determining the magnitude of antigen-induced proliferative responses. These data are consistent with the hypothesis that cell traffic after sensitization plays an important role in determining the immune responsiveness of lymph nodes.

Animals

Distinct contributions of Aire and antigen-presenting-cell subsets to the generation of self-tolerance in the thymus.

The contribution of thymic antigen-presenting-cell (APC) subsets in selecting a self-tolerant T cell population remains unclear. We show that bone marrow (BM) APCs and medullary thymic epithelial cells (mTECs) played nonoverlapping roles in shaping the T cell receptor (TCR) repertoire by deletion and regulatory T (Treg) cell selection of distinct TCRs. Aire, which induces tissue-specific antigen expression in mTECs, affected the TCR repertoire in a manner distinct from mTEC presentation. Approximately half of Aire-dependent deletion or Treg cell selection utilized a pathway dependent on antigen presentation by BM APCs. Batf3-dependent CD8α⁺ dendritic cells (DCs) were the crucial BM APCs for Treg cell selection via this pathway, showing enhanced ability to present antigens from stromal cells. These results demonstrate the division of function between thymic APCs in shaping the self-tolerant TCR repertoire and reveal an unappreciated cooperation between mTECs and CD8α⁺ DCs for presentation of Aire-induced self-antigens to developing thymocytes.

Animals

Impairment of antigen-presenting cell function by ultraviolet radiation.

UV light irradiation of BALB/c mice was found to result in impairment of antigen-presenting cell function. Adherent trinitrophenyl-derivatized cells from the peritoneal exudate cell population or the spleen of UV-treated donors could not induce hapten-specific delayed hypersensitivity responses in UV-irradiated syngeneic mice, whereas adherent trinitrophenyl-derivatized cells from normal mice were able to do so. The failure to induce immunity in UV-treated mice by utilizing UV-treated adherent antigen-presenting cells was associated with the development of antigen-specific suppressor T cells. The implication of these results for UV-induced carcinogenesis is discussed.

Animals

The removal of "nucleic acid" from an antigen present in the livers of chickens with erythroblastosis.

The recovery of viral antigen in a pure form from tissue can be hindered by the small amount of antigen present relative to the remaining normal tissue constituents with which the antigen may form loose associations. An antigen from the livers of chickens with erythroblastosis had earlier seemed to be associated with nucleic acid. The present work suggests that this material, which reacts with reagents for DNA and RNA but is soluble in 0.2N HClO4, is only a contaminant and not an integral part required for complete activity of the antigen. Treatment with 0.2N HClO4, together with precipitation of the active protein by 60% saturation of aqueous solutions with ammonium sulphate, removes the nucleic acid without destroying antigenicity. The procedure results in a marked degree of purification of the antigen (732-fold with respect to original liver protein) but contaminants still remain. A contaminant absorbing at 260 nm resisting extraction with 0.2N HClO4, can be partly eliminated if the solution is treated with ether-alcohol to remove lipid.

Animals

Local delivery of interferon restores antigen presentation and sensitizes medulloblastoma to T cell killing.

Medulloblastomas are considered immunologically cold and refractory to immunotherapy. One factor contributing to their low immunogenicity is impaired antigen presentation, which allows tumor cells to evade cytotoxic T cells. Using a syngeneic mouse model of medulloblastoma, we demonstrate that despite low expression of MHC class I on tumor cells, depletion of CD8+ T cells accelerates tumor growth, whereas adoptive transfer of tumor-reactive CD8+ T cells prolongs survival. These antitumor effects rely on T cell-derived interferon gamma (IFNγ), which induces MHC class I on tumor cells and facilitates tumor cell killing by T cells. Importantly, delivering IFNγ directly into tumors via convection-enhanced delivery enhances CD8+ T cell-mediated killing of tumor cells and significantly prolongs survival in tumor-bearing mice. These studies highlight the importance of T cells in controlling brain tumors and the value of IFNγ as an adjuvant for T cell-based immunotherapy.

Animals

Immunological tolerance: high-dose antigen-induced suppressor cells from tolerant animals inactivate antigen-presenting macrophages.

Studies were carried out to characterize the target cell for the activity of suppressor cells induced in highzone tolerance to deaggregated human gamma globulin (HGG). We applied an in vitro system for the initiation of an immune response, consisting of culturing spleen lymphocytes on HGG-fed macrophages, in which initiator T cells are generated. These cells, when injected into the foot pads of syngeneic mice, recruit specific anti-HGG effector T lymphocytes. We found that HGG-fed macrophages were incapable of signaling spleen cells from HGG-tolerant animals to generate initiator cells. Spleen cells from tolerant animals, when mixed with spleen cells from normal donors, inhibited the capacity of the normal population to give rise to initiator cells after culture on HGG-fed macrophages. Thus, suppressor T cells, which inhibit education of T cells by antigen-fed macrophages, exist in the tolerant spleen. Spleen cells from HGG-tolerant animals, when seeded on macrophages fed simultaneously with HGG and keyhole limpet hemocyanin (KLH), also prevented the macrophages from signaling an anti-KLH response. Spleen cells from HGG-tolerant animals from which the suppressor cells were depleted by "affinity chromatography" on histamine columns, when seeded on macrophages fed with HGG and KLH, generated initiators to both antigens. It appears, therefore, that suppressor cells act at the level of antigen-presenting macrophages, affecting macrophages fed with the tolerogen, and therefore affecting also the immunogenic effect of other antigens presented by the same macrophages. By testing the mode of action of suppressor cells on the tolerogen-fed macrophage, we found that the suppressors manifest a cytotoxic effect on such macrophages. We propose that the suppressor cell is, in fact, an anti modified-self killer, acting on macrophages possessing surface self-antigens "modified" by the tolerogen. The similarity in cell-surface markers between suppressors and anti modified-self killers supports this concept.

Animals

RENBP inhibition amplifies metabolic glycan labeling efficiency of antigen-presenting cells in vitro and in vivo.

Metabolic glycoengineering of unnatural sugars provides a powerful tool to introduce unique chemical tags onto cell membrane for subsequent conjugation of cargos. However, the metabolic glycan labeling efficiency of antigen-presenting cells (APCs), the key mediators of adaptive immunity, is often low. Here, we report that APCs upregulate GlcNAc 2-epimerase (RENBP) and that RENBP inhibition leads to improved labeling efficiency of tetraacetyl-N-azidoacetylmannosamine (AAM) in APCs, including dendritic cells (1.2-fold), macrophages (1.3-fold), and B cells (1.4-fold) in vitro. RENBP inhibition can preferentially enhance AAM labeling efficiency in APCs than in non-APCs and selectively enhance the labeling efficiency of AAM over azido-galactosamine. We further demonstrate that RENBP inhibitors can improve AAM-mediated labeling of B cells and other APCs in vivo, with the largest enhancement for B cells (>3-fold) for 7 days. Our study uncovers a facile approach to improving metabolic glycan labeling of APCs, enabling the development of APC-targeted immunotherapies.

Animals

Antigen presentation as a factor in the protective immune response to renal infection.

Host protection against renal infection may be augmented by active immunization against the causative organism. In these experiments we have investigated the effect of varying amounts and methods of presentation of bacterial antigen on the seconday immune response. Primary immunization with varying amounts of both killed and live antigen did not affect the nature of the secondary immune response although active renal infection did have a noticeable effect on the titre of serum antibody during the primary immune response. The experiments confirmed the presence of immunological memory to the somatic antigen of E. coli and showed that memory persisted for at least 6 months after primary immunization. Experiments have also been carried out which have demonstrated that memory to the somatic antigen of E. coli is carried by the B lymphocyte.

Animals

Intratumoral Sotigalimab with Pembrolizumab Induces Rapid Activation of Antigen-Presenting Cells and Drives Antitumor Responses in Non-injected Tumors in Metastatic Melanoma: A Phase I/II Study.

UNLABELLED: Immune checkpoint blockers (ICB) improve outcomes in metastatic melanoma (MM), but resistance limits benefit. This phase I/II (NCT02706353) study evaluated intratumoral sotigalimab (anti-CD40 agonist) with pembrolizumab in 32 patients with ICB-naïve MM. Primary endpoints were safety and objective response rate (ORR). Sotigalimab was well tolerated. At the recommended phase II dose, the ORR was 50%, and the disease control rate was 92%, with ORRs of 67% in injected and 50% in non-injected tumors. Multiomic analyses of tumor and blood showed that sotigalimab effectively engaged the CD40 pathway, boosting infiltration and activation of myeloid cells, including CD11c+DC-LAMP+ dendritic cells and macrophages. The combination therapy activated innate and adaptive immunity in injected tumors and cytotoxic responses in non-injected tumors. T-cell receptor sequencing showed increased T-cell clonality with expanded new clones shared across tumors. Clinical responses correlated with these immunologic changes but not with baseline features associated with response to anti-PD-1 monotherapy. SIGNIFICANCE: In this study, we provide compelling data that intratumoral sotigalimab combined with pembrolizumab is safe, activates antigen-presenting cells, and elicits broad innate and adaptive immune responses in both injected and non-injected tumors, supporting further randomized phase II trials to evaluate sotigalimab's potential to enhance anti-PD-1 therapy through "in situ" immunization.

Humans

A new particulate antigen present in serum.

A new antigen has been encountered in the serum of 10 blood donors. It is distinct from Australia antigen, consists of spherical particles 21-23 nm in diameter and the corresponding antibody is widely distributed in the population.

Adolescent

Hybrid myelomas producing antibodies against a human neuroblastoma antigen present on fetal brain.

Spleen cells from mice immunized with a cultured human neuroblastoma were hybridized with the mouse plasmacytoma P3X63Ag8. Hybrid myelomas were screened for production of antibodies that reacted with human neuroblastomas but not with cells from other tissues. One of these hybridoma antibodies reacted with an antigen present on the six human neuroblastomas tested, one of two retinoblastomas, a glioblastoma, and fetal brain, but did not react with other tumors or tissues including adult human brain.

Antigens, Neoplasm

The penicillin antigens: presentation in the leucocyte migration test (a study with penicillin and penicilloylated proteins).

The response of the leucocyte migration test (LMT) to benzyl penicillin (BP) was compared with the response of LMT to benzyl penicilloylated human gamma globulins (BPO HGG) and to benzyl penicilloylated human serum albumin (BPO HSA). Patients with and without hypersensitivity to penicillins were investigated. No statistical differences were found between the response to BP and to BPO HGG or BPO HSA; this finding supports the fact that the BPO groups generated by BP are mainly implicated in the triggering of the LMT when BP is used as antigen. A strongly penicilloylated HSA appeared to be the most effective mode of the antigen presentation for the detection of BPO-specific cellular immunity in patients with and without clinical symptoms of hypersensitivity to the penicillins.

Adult