Production of migration inhibitory factor by Listeria-immune mouse T lymphocytes, but not B lymphocytes

Antigens and B cell mitogens have been reported to induce migration inhibition factor (MIF) production by mouse B cells. Immune resistance to the intracellular bacterium, Listeria monocytogenes is thought to involve T cells, but not B cells. Since Listeria-derived components are B cell, but not T cell mitogens, it was important to determine whether these materials could stimulate secretion of the lymphokine, MIF by T cells, B cells, or both. Thus populations of whole, unfractionated spleen cells, obtained from normal and Listeria-immune BDF1 mice, were cultured with or without 100 micrograms/ml of Listeria intracellular product (LIP). The culture supernatants obtained 24 h later were assayed for MIF activity using the in vitro macrophage migration inhibition assay. Data obtained show that immune T lymphocytes release MIF in response to specific Listeria antigens, but that spleen B cells from immune and normal mice, obtained as immune, nylon-wool-adherent cells treated with anti-T-cell serum plus complement, are not capable of releasing MIF. This suggests that release of lymphokines by Listeria-immune or normal B cells stimulated with Listeria-derived antigens and mitogens is unlikely to contribute to resistance against Listeria in vivo.     

A factor present in human milk, but not colostrum, which is cytotoxic for human lymphocytes

This study reports the presence of a factor in human milk which is cytotoxic to both autologous and heterologous human peripheral blood mononuclear cells (PBMC). The cytotoxic effect was measured by the release of 51Cr from labelled cells or by the inability of PBMC to exclude trypan blue following exposure to milk. The cytotoxic factor was not dialysable or sensitive to heating at 56 degrees C for 30 min, and could not be adsorbed from milk by PBMC. It lysed lymphocytes harvested from colostrum and from autologous or heterologous milk. The cytotoxic factor in the milk was different from the factor found in colostrum which inhibits the proliferation of mitogen stimulated PBMC, but which is not cytotoxic. Testing of serial samples of milk from the same mother indicated that the factor could be detected in the milk after 3-4 days post partum. We conclude that factors cytotoxic for human PBMC appear in human milk early in lactation.     

Interleukin 5 is a differentiation factor for cytotoxic T lymphocytes

The role of IL-5 on the generation of cytotoxic T lymphocytes (CTL) was analysed using a culture system in which production of T helper cell factors was abrogated by exposure of the stimulator cells to ultraviolet irradiation. Supernatants from a T helper cell line (2.19 sup), recombinant (r) IL-2, rIL-4 and rIL-5 were then tested for the capacity to replace T cell help, on the generation of CTL. The results showed that the specific CTL response, in unseparated spleen cells, could be reconstituted by either 2.19 sup, IL-2 or IL-4. However, if the responder cells were purified in nylon wool, only 2.19 sup or rIL-4 plus rIL-5, but not each lymphokine per se, reconstituted the CTL response. Because IL-5 does not support T cell proliferation, it is suggested that IL-5 induces differentiation of immature precursors into CTL. Based on these findings and in an attempt to conciliate the conflicting views that have emerged from different reports, as to whether IL-2 by itself could support generation of CTL in purified T cells, a hypothesis is formulated, suggesting that T cell at different stages of differentiation require distinct lymphokines to acquire CTL effector function.     

T cell-derived tumour necrosis factor is essential, but not sufficient, for protection against Mycobacterium tuberculosis infection

Tumour necrosis factor (TNF) is critical for sustained protective immunity against Mycobacterium tuberculosis infection. To investigate the relative contributions of macrophage- and T cell-derived TNF towards this immunity T cells from wild-type (WT) or TNF-/- mice were transferred into RAG-/- or TNF-/- mice which were then infected with M. tuberculosis. Infected RAG-/- mice and RAG-/- recipients of TNF deficient T cells developed overwhelming infection, with extensive pulmonary and hepatic necrosis and succumbed with a median of only 16 days infection. By contrast, RAG-/- recipients of WT T cells showed a significant increase in survival with a median of 32 days. Although initial bacterial growth was similar in all groups of RAG-/- mice, the transfer of WT, but not TNF-/-, T cells led to the formation of discrete foci of leucocytes and macrophages and delayed the development of necrotizing pathology. To determine requirements for macrophage-derived TNF, WT or TNF-/- T cells were transferred into TNF-/- mice at the time of M. tuberculosis infection. Transfer of WT T cells significantly prolonged survival and reduced the early tissue necrosis evident in the TNF-/- mice, however, these mice eventually succumbed indicating that T cell-derived TNF alone is insufficient to control the infection. Therefore, both T cell- and macrophage-derived TNF play distinct roles in orchestrating the protective inflammatory response and enhancing survival during M. tuberculosis infection.     

gamma-Glutamyltransferase is a Helicobacter pylori virulence factor but is not essential for colonization

The contribution of glutamyl transpeptidase (GGT) (gamma-glutamyltransferase [EC 2. 3. 2. 2]) to Helicobacter pylori virulence was investigated in piglets and mice using GGT-deficient isogenic strains. All animals became colonized. However, the bacterial load was significantly lower for mutant bacteria than for parent strains. These results suggest that GGT activity provides an advantage to H. pylori in colonization.     

Interleukin-6 enhances human Ig production,but not as a terminal differentiation factor for B lymphocytes

Most B-cell differentiation systems are complicated by the fact that they are both T-cell- and monocyte-dependent. Immobilized anti-CD3 antibodies induce monocyte-independent T-cell activation, allowing investigation of the role of interleukin-6 (IL6) in the process of B-cell differentiation. We observed that in this system, the addition of monocytes to purified lymphocytes does not influence T-cell proliferation but it does enhance the induction of Ig production. IL6 can specifically replace monocytes in this enhancing effect on both IgM and IgG production. Anti-CD3-induced Ig production appears to be dependent on both IL2 and IL6 since it was inhibited by anti-CD25 (anti-IL2-R) antibodies as well as by anti-IL6 antibodies. Kinetic studies of IL6 addition showed that IL6 is only necessary during the first two days of culture. Our data indicate that IL6 plays an essential role in anti-CD3-induced Ig production, but not as a terminal differentiation factor.     

Interleukin-6 enhances human Ig production, but not as a terminal differentiation factor for B lymphocytes

Most B-cell differentiation systems are complicated by the fact that they are both T-cell- and monocyte-dependent. Immobilized anti-CD3 antibodies induce monocyte-independent T-cell activation, allowing investigation of the role of interleukin-6 (IL6) in the process of B-cell differentiation. We observed that in this system, the addition of monocytes to purified lymphocytes does not influence T-cell proliferation but it does enhance the induction of Ig production. IL6 can specifically replace monocytes in this enhancing effect on both IgM and IgG production. Anti-CD3-induced Ig production appears to be dependent on both IL2 and IL6 since it was inhibited by anti-CD25 (anti-IL2-R) antibodies as well as by anti-IL6 antibodies. Kinetic studies of IL6 addition showed that IL6 is only necessary during the first two days of culture. Our data indicate that IL6 plays an essential role in anti-CD3-induced Ig production, but not as a terminal differentiation factor.     

Cytotoxic T lymphocytes, chemokines and antiviral immunity.

Evidence that CD8+ CTLs produce chemokines following engagement of viral antigens, and that MIP-1alpha is required for an inflammatory response to virus challenge, suggests that these molecules are key elements in the generation of effective antiviral immunity. Here, David Price and colleagues argue that the antigen-dependent release of chemokines by CTLs provides an elegant mechanism linking localization, amplification and coordination of the antiviral immune response to specific recognition of infected host cells beyond the confines of the lymphoid system.     

Chondroblastoma is an osteoid-forming, but not cartilage-forming neoplasm

Chondroblastoma is defined as a 'benign tumour, characterized by highly cellular and relatively undifferentiated tissue composed of rounded or polygonal chondroblast-like cells' and the 'presence of cartilaginous intercellular matrix' (WHO). An extensive analysis of the extracellular matrix composition and gene expression pattern of a large series of chondroblastoma cases shows, however, that type II collagen, which is the main component of any cartilage matrix, is not expressed by the neoplastic cells of this tumour entity and is not deposited into the extracellular tumour matrix. Instead, osteoid and fibrous matrix is formed, with its typical biochemical composition. The multifocal expression of aggrecan proteoglycan in most chondroblastomas explains the bluish, pseudo-chondroid appearance of some of the matrix-rich areas of chondroblastomas. This study did not show chondroid matrix formation or chondroblastic cell differentiation in chondroblastomas, suggesting that chondroblastoma should be classified as a specific bone-forming, rather than cartilage-forming neoplasm.     

Cytotoxic T lymphocytes recognize and lyse chondrocytes under inflammatory,but not non-inflammatory conditions

The human major histocompatibility complex (MHC) class I allele HLA-B27 is strongly associated with seronegative spondyloarthropathies including ankylosing spondylitis and reactive arthritis. Although of unknown aetiology, one hypothesis suggests that a cytotoxic T cell (CTL) response against a self-antigen at sites of inflammation, such as entheses or joints may be involved. The chondrocyte is one of the major specialized cell types found both in articular cartilage and cartilaginous entheses and therefore is a possible source of such an antigen. CTL recognition of these cells is a potential mechanism for inflammation and cartilage damage, both through direct lysis of chondrocytes and the secretion of pro-inflammatory cytokines such as tumour necrosis factor and interferon-gamma (IFN-gamma). We test the feasibility of this hypothesis by examining the ability of chondrocytes to present antigen to CTL in vitro. Chondrocytes isolated from the ribcages of mice did not constitutively express detectable levels of MHC class I by fluorescence-activated cell sorting analysis. In addition, they were resistant to lysis by alloreactive and influenza A virus nucleoprotein (NP)-specific CTL. However, treatment of chondrocytes with IFN-gamma up-regulated MHC class I expression and rendered the cells susceptible to lysis by CTL. Similarly, IFN-gamma-treated chondrocytes infected with influenza A virus were recognized by NP-specific CTL, though with variable efficiency. Thus, we suggest that under certain circumstances CTL-mediated lysis of chondrocytes is potentially a potent mechanism for cartilage damage in vivo, but that low levels of MHC class I on healthy chondrocytes protects from immune recognition in health.     

TCGF III/P40 is produced by naive murine CD4+ T cells but is not a general T cell growth factor

Several antigen-specific T cell lines were found to secrete a lymphokine upon activation by antigen or lectin that was provisionally termed T cell growth factor III (TCGF III) because it induced the proliferation of a CD4+ T cell clone independently from IL2 and IL4. Amino acid sequence analysis (and the functional properties of TCGF III) revealed that TCGF III was identical with a recently identified lymphokine termed P40. TCGF III/P40 was not only produced by long-term cultured T cell lines but also upon stimulation of freshly isolated Mlsa-reactive T cells. In addition, naive CD4+ T cells secreted TCGF III/P40 upon activation by lectin or allo-major histocompatibility complex structures. However, in spite of its growth-promoting activity for a CD4+ T cell clone this lymphokine does not appear to function as a general growth factor for T cells.     

Rifampicin quinone is an immunosuppressant, but not rifampicin itself

Rifampicin as a potential immunosuppressive agent was tested in the rat. A freshly made-up solution of this labile antibiotic in a daily dosage of 20 mg/kg iv did not affect mean graft survival time in a heterotopic heart transplant model. However, when stored solutions of rifampicin were used mean graft survival time was significantly prolonged (from 8.2 +/- 0.4 days with the controls to 18.1 +/- 1.2 days). A similar prolongation was observed when rifampicin quinone, the major rifampicin oxidation product, was administered. We conclude that the immunosuppressive effect ascribed to rifampicin is in fact due to its oxidation product, rifampicin quinone.     

Mitogenic effect of beryllium sulfate on mouse B lymphocytes but not T lymphocytes in vitro

Beryllium metal and its salts can produce disease in man and in animal models. Beryllium disease is thought to involve cell-mediated immunity and an antigen-dependent response by beryllium-specific T cells. Beryllium salts have been shown to stimulate lymphocyte proliferation and release of lymphokines, and to induce granuloma formation and delayed-type hypersensitivity reactions in mice, guinea pigs and man. The studies described here were designed to test the hypothesis that a second lymphocyte population, B cells, may be responding nonspecifically to beryllium. Different populations of BDF1 mouse lymphocytes were cultured in the presence of varying concentrations of beryllium sulfate (BeSO4), and the increase in 125-iodouracildeoxyriboside uptake after 72 h in culture was determined. The data show that BeSO4 is weakly mitogenic for normal mouse spleen cells. Furthermore, BeSO4 is mitogenic for normal and nude mouse spleen B cells and not for spleen T cells or thymocytes in vitro. These findings suggest that BeSO4 can stimulate B cells nonspecifically, and support the hypothesis that polyclonal activation of B cells by beryllium may occur.     

Transduction efficiency of MLV but not of HIV-1 vectors is pseudotype dependent on human primary T lymphocytes

The success of several gene therapeutic approaches requires efficient transduction of human primary T lymphocytes. For this it is important to enhance the transduction efficiency, and this can be achieved by various means, mainly technical development of transduction procedures and use of different vectors and vector pseudotypes. We analyzed the transduction efficiency of an HIV-1 vector encoding enhanced green fluorescent protein (GFP) as a marker gene and pseudotyped with the envelopes of MLV-A, MLV-10A1, GaLV, RD114, and VSV for human primary T lymphocytes in comparison to an MLV vector pseudotyped with the same envelopes. Pseudotyping of the MLV vector with the envelopes of 10A1 and GaLV resulted in efficient transduction of preactivated human primary T lymphocytes (32.4% and 32.7% CD3⁺/GFP⁺ cells, respectively) while MLV-A (14.0%), RD114 (8.8%), and VSV (1.5%) envelopes were less efficient when using titrated vector stocks equilibrated to a multiplicity of infection of 1. In contrast, the HIV-1 vectors pseudotyped with these envelope proteins transduced preactivated T lymphocytes with similar efficiency (approx. 20% CD3⁺/GFP⁺ cells). Thereby, CD4⁺ and CD8⁺ T lymphocyte subpopulations were transduced at equivalent levels. The similar performance of the different HIV-1 vector pseudotypes may be due in part to the similar half-lives of the vector particles. Independently of the envelope used for pseudotyping neither the MLV nor the HIV-1 vectors yielded any significant transduction in nonactivated T lymphocytes (below 0.55% of GFP⁺ cells)Abbreviations FCS Fetal calf serum - GaLV Gibbon ape leukemia virus - GFP Green fluorescent protein - GFU Green fluorescent colony-forming unit - GP Glycoprotein - HIV Human immunodeficiency virus - IL Interleukin - mAb Monoclonal antibody - MLV Murine leukemia virus - MOI Multiplicity of infection - PBL Peripheral blood lymphocyte - PHA Phytohemagglutinin - VSV-G Vesicular stomatitis virus glycoprotein     

The shared tumor-specific antigen encoded by mouse gene P1A is a target not only for cytolytic T lymphocytes but also for tumor rejection

A number of human tumor antigens have been characterized recently using cytolytic T lymphocytes (CTL) as screening tools. Some of them are encoded by MAGE-type genes, which are silent in normal tissues except in male germ cells, but are activated in a variety of tumors. These tumor-specific shared antigens appear to be promising targets for cancer immunotherapy. However, the choice of these antigens as targets has been questioned because of the lack of direct evidence that in vivo responses against such antigens can lead to tumor rejection. The antigen encoded by the mouse gene P1A represents the only available animal model system for MAGE-type tumor antigens. We show here that mice immunized by injection of L1210 leukemia cells expressing P1A and B7-1 (L1210.P1A.B7-1) are efficiently protected against a challenge with a lethal dose of mastocytoma P815 tumor cells, which express P1A. Mice immunized with L1210 cells expressing B7-1 but not P1A were not protected. Furthermore, we observed that P1A-transgenic mice, which are tolerant to P1A, were not protected after immunization with L1210.P1A.B7-1. These results demonstrate that the immune response to P1A is the major component of the tumor rejection response observed in normal mice, and support the use of tumor-specific shared antigens as targets for the immunotherapy of human cancer.     

Socioeconomic status is a risk factor for allergy in parents but not in their children

BACKGROUND: Allergic diseases are more prevalent in affluent countries, which has been attributed to life-style factors. Life-style habits may also differ between socioeconomic (SES) classes. The objective of this paper therefore was to evaluate if SES had an impact on the development of atopic disorders. METHODS: A total of 1314 German children were followed-up in an observational birth cohort study to 6 years of age. Parents filled in questionnaires, and had multi-allergen screening tests for sensitization. Indoor allergen concentrations were determined by ELISA. Children were examined regularly up to 6 years, specific serum IgE values were determined by CAP-Rast-Feia. RESULTS: The risk of aeroallergen sensitization (odds ratio 1.76; 95% CI 1.30-2.37), and the lifetime prevalence of hay fever (2.36; 1.76-3.17), and asthma (1.74; 1.08-2.80), but not of atopic dermatitis (AD: 0.90; 0. 54-1.51) was elevated in parents of high compared to low SES. With high SES the risk of smoking in pregnancy (0.35; 0.23-0.51), in the home (0.31; 0.21-0.46), pet ownership (0.37; 0.26-0.55), high mite (0.42; 0.25-0.74), and high cat (0.38; 0.18-0.82) allergen concentration in house dust was reduced, but elevated for breastfeeding over more than 6 months (4.67; 2.9-7.48). In children, even after controlling for other risk factors, only the risk of AD from 3 to 6 years (2.42; 1.42-4.14) was elevated in families with high SES, but not of AD in infancy or of any other atopic disorder. CONCLUSIONS: While parents of high SES have a higher prevalence of inhalative allergies, their favourable life-style prevents the development of atopic disorders in their children, except for AD beyond infancy.