The in vivo antiviral activity of interleukin-12 is mediated by gamma interferon.

The injection of 20 ng of mouse interleukin-12 (IL-12) protects mice from a lethal infection with encephalomyocarditis virus. In vitro, an anti-gamma interferon (anti-IFN-gamma) monoclonal antibody but not an anti-IL-12 monoclonal antibody neutralizes the antiviral activity present in the supernatants of splenocytes stimulated with IL-12. Finally, IL-12 fails to protect 129 Sv/Ev IFN-gamma R0/0 mice against encephalomyocarditis virus infection. These results demonstrate that IL-12 exerts its antiviral activity through the induction of endogenous IFN-gamma.     

Enhancement of in vivo cell-mediated immune responses by three distinct cytokines

The ability of recombinant/purified cytokines to augment delayed-type hypersensitivity (DTH) responses was investigated. Suboptimal doses of haptenized SC were treated in vitro with purified or recombinant derived cytokines and tested for their ability to enhance DTH in vivo. With the use of this protocol, it was shown that both human and mouse rIL-6, as well as mouse rTNF-alpha, potentiated DTH in a dose-dependent manner. In accordance with these data, IL-6/TNF-alpha-containing supernatant from long term nonlymphoid cell lines also possessed the ability to augment DTH. By using the same protocol, we have also identified T cell hybridomas that produce DTH-augmenting activity constitutively. The hybridoma-derived factor, termed the T cell enhancing factor (TCEF), was functionally distinguishable from the defined cytokines IL-1 through IL-6, IFN-gamma, and TNF by bioassay. Furthermore, RNA derived from the hybridoma failed to hybridize with cDNA probes specific for IL-1 to IL-6, IFN-gamma, TNF-alpha, and granulocyte-macrophage CSF. Further characterization of the serum-free conditioned media derived from the hybridoma indicated that the TCEF was a soluble acid labile glycoprotein (Mr greater than 30,000). Finally, we investigated the cellular requirements for DTH augmentation by IL-6, TNF-alpha, and TCEF; all are dependent upon the presence of T cells in the immunizing inoculum. We propose that these cytokines play a critical role in the development of DTH responses in vivo.     

Spectrum of in vivo and in vitro cell-mediated immune responses in coccidioidomycosis.

The cell-mediated immune responses of 12 healthy, coccidioidin skin-test positive subjects (Group I) were compared with those of 15 healthy, coccidioidin skin-test positive persons who had primary asymptomatic coccidiodomycosis, (Group II), 12 patients with active, pulmonary coccidioidomycosis (Group III), four patients with disseminated disease (Group IV), and five patients who had been in clinical remission for 1 year or longer (Group V). Lymphocytes from healthy subjects in Groups I and II responded in vitro to Coccidioides immitis antigen by undergoing an increased DNA synthesis (lymphocyte transformation) and/or by producing macrophage migration inhibitory factor (MIF). In contrast, patients in Groups III and IV failed to respond to Coccidioides antigens in vivo (skin tests) or in vitro (lymphocyte transformation and production of MIF). The responses of subjects in Group V with inactive disease fell in between those of healthy donors in Groups I and II and patients in Groups III and IV. The cellular immune defect, in terms of antigen recognition, appeared to be specific for C. immitis in all but one patient.     

Survival strategies of Entamoeba histolytica: Modulation of cell-mediated immune responses

Tissue invasion and disease associated with the protozoan Entamoeba histolytica has long been connected with suppression of host cellular immunity. Dampening of the host's defences may facilitate survival of amoebae in extraintestinal sites and development of the characteristic amoebic abscesses. In recent years, several studies have begun to clarify, at the cellular level, the specific effects E. histolytica has on immune cell accessory and effector cell functions. Here, Darren Campbell and Kris Chadee discuss the parasite's multiple modulatory effects on macrophages and T cells and how this manipulation of immune defences may enable the parasite to remain viable in the host. They suggest the putative amoebic molecules involved and potential modulation by the cytokines: interleukins IL-4 and IL-10 and transforming growth factor-beta.     

The antiviral response to gamma interferon

A role for alpha/beta interferon (IFN-alpha/beta) in the IFN-gamma antiviral response has long been suggested. Accordingly, possible roles for autocrine or double-stranded-RNA (dsRNA)-induced IFN-alpha/beta in the IFN-gamma response were investigated. Use was made of wild-type and a variety of mutant human fibrosarcoma cell lines, including mutant U5A cells, which lack a functional IFN-alpha/beta receptor and hence an IFN-alpha/beta response. IFN-gamma did not induce detectable levels of IFN-alpha/beta in any of the cell lines, nor was the IFN-gamma response per se dependent on autocrine IFN-alpha/beta. On the other hand, a number of responses to dsRNA [poly(I). poly(C)] and encephalomyocarditis virus were greatly enhanced by IFN-gamma pretreatment (priming) of wild-type cells or of mutant cells lacking an IFN-alpha/beta response; these include the primary induction of dsRNA-inducible mRNAs, including IFN-beta mRNA, and, to a lesser extent, the dsRNA-mediated activation of the p38 mitogen-activated protein (MAP) kinase(s). IFN-gamma priming of mRNA induction by dsRNA is dependent on JAK1 and shows biphasic kinetics, with an initial rapid (<30-min) response being followed by a more substantial effect on overnight incubation. The IFN-gamma-primed dsRNA responses appear to be subject to modulation through the p38, phosphatidylinositol 3-kinase, and ERK1/ERK2 MAP kinase pathways. It can be concluded that despite efficient priming of IFN-beta production, the IFN-alpha/beta pathways play no significant role in the primary IFN-gamma antiviral response in these cell-virus systems. The observed IFN-gamma priming of dsRNA responses, on the other hand, will likely play a significant role in combating virus infection in vivo.     

Modulation of immune responses by suppressor T cells.

The activity of suppressor T cells has been demonstrated in almost every phase of the immune response. These regulatory cells modulate both humoral and cell-mediated immunity utilizing antigen-specific and nonspecific mechanisms. For comparative purposes two murine models are described, the nonspecific suppressor T cell stimulated by the mitogen concanavalin A and the antigen-specific suppressor T cell stimulated by injection of the synthetic terpolymer acid 60-L-alanine30-L-tyrosine10 (GAT) in nonresponder mice. These two T cells are similar to expression of Ly alloantigens, ability to inhibit antibody responses, and the mediation of suppression, at least in part, by soluble products. However, differences in radio-resistance and antigenic specificity of the suppressor T cells, as well as differences in molecular characteristics of the soluble factors and their targets suggest that these T cells regulate the immune response by different mechanisms. The relationship of these two suppressor T cells to other nonspecific and antigen-specific suppressor T cells is discussed.     

Influences of roxithromycin on cell-mediated immune responses.

We investigated the effects of roxithromycin (RXM), a synthesized macrolide antibiotic on murine cellular immune responses by examining the in vitro proliferative response of lymphocytes, interleukin 1 (IL-1) production and interleukin 2 (IL-2) production. RXM was orally administered to BALB/c mice at a dose of 5 mg/kg once a day for 42 days. Spontaneous blastic activity of lymphocytes prepared from mice administered with RXM for 7 days was higher than those from control mice. The activity peaked at the 14th day, and then decreased gradually to control levels by the 42nd day. Time kinetics of lymphocyte blastogenesis to concanavalin A showed a pattern similar to that observed in spontaneous blastic activity. Oral administration of RXM also influenced cytokine production; short-term (for 14 days) administration of RXM enhanced both IL-1 and IL-2 production but long-term (for 42 days) administration inhibited them.     

Interleukin 2 in cell-mediated immune responses

The lymphokine Interleukin 2 (IL2) restores T cell responses in a number of in vitro systems where immunogenicity has been compromised. UV irradiation of the stimulating allogeneic cells in a mixed leukocyte culture eliminates the production of cytotoxic T lymphocytes and greatly reduces the DNA synthesis response. IL2 restores both parameters. UV-irradiated stimulators are also unable to induce the normal production of IL2 which is observed in a mixed leukocyte culture. The cytotoxic activity of allogeneically stimulated thymocytes is almost completely lost within 24 hours after removal of IL2 at 5 days, indicating that the lymphokine is continuously required to maintain CTL. Thymocytes in 4-day cultures do not adsorb IL2 unless they are simultaneously activated with a mitogen. Finally, IL2 does not adequately restore a secondary response to the purified protein derivative of tuberculin (PPD) in adherent-cell-depleted cultures, indicating that macrophages, in addition to being required for IL2 production, have other functions. These probably include the presentation of soluble antigens to responding cells.     

Evasion from NK cell-mediated immune responses by HIV-1

Human immunodeficiency virus type 1 (HIV-1) mostly owes its success to its ability to evade host immune responses. Understanding viral immune escape mechanisms is a prerequisite to improve future HIV-1 vaccine design. This review focuses on the strategies that HIV-1 has evolved to evade recognition by natural killer (NK) cells.     

Modulation of humoral immune responses by Tinospora malabarica in experimental animals.

Effect of alkaloidal fraction of aqueous extract of T. malabarica (Tm) was studied on humoral antibody responses in rats and guineapigs. The anti-SRBC haemagglutination titre was found to be enhanced in rats pretreated with Tm (2.5 mg/kg). Passive cutaneous anaphylaxis (PCA) in rats was also increased in Tm treated group. In vitro experiments with sensitized rat peritoneal mast cells showed a significant decrease in antigen-induced various spasmogens on isolated guineapig ileum.     

Modulation of host immune responses by nematode cystatins

Parasitic nematodes, living in the intestinal tract or within tissues of theirs hosts, are constantly exposed to an array of immune effector mechanisms. One strategy to cope with the immune response is the release of immunomodulatory components that block effector mechanisms or interact with the cytokine network. Among the secreted nematode immunomodulators, cysteine protease inhibitors (cystatins) are shown to be of major importance. Nematode cystatins inhibit, among others, proteases involved in antigen processing and presentation, which leads to a reduction of T cell responses. At the same time nematode cystatins modulate cytokine responses, the most prominent trait being the upregulation of IL-10, a Th2 cytokine, by macrophages. In this situation, IL-10 leads among others to downregulation of costimulatory surface molecules of macrophages. These properties contribute to induction of an anti-inflammatory environment, concomitant with a strong inhibition of cellular proliferation. This setting is believed to favour the survival of worms. An opposite activity of nematode cystatins is the upregulation of production of inducible nitric oxide by IFN-gamma activated macrophages, an intrinsic property of natural cysteine protease inhibitors. This shows that these proteins can act as proinflammatory molecules under certain circumstances. A comparison of the immunomodulatory effects of cystatins of filarial nematodes with homologous proteins of the free-living nematode Caenorhabditis elegans revealed distinct differences. Caenorhabditis elegans cystatins induce the production of the Th1 cytokine IL-12, in contrast to filarial cystatins that upregulate IL-10. Caenorhabditis elegans cystatins hardly inhibit cellular proliferation. These data suggest that cystatins of parasitic nematodes have multiple, specific capacities for immunomodulation, acting in parallel on different immune effector mechanisms. Elucidation of the mechanisms involved might be useful in the development of immunotherapeutic reagents in the future.     

Modulation of expression of the human gamma interferon gene in E. coli by site-directed mutagenesis

Plasmids expressing 2 forms of human immune interferon (IFN-gamma) in E. coli have been constructed: 1) pIFNTacI which expresses IFN-gamma with an N-terminal amino acid sequence of met-cys-tyr-cys-gln-, and 2) pIFNTacII which is a derivative of pIFNTacI from which the 9 base pairs (bp) coding for the cys-tyr-cys have been deleted. Quantitation of Western blots showed that approximately 10-fold more IFN-gamma was produced in cells harboring pIFNTacII (7.5% of total cellular protein) as compared to pIFNTacI. The IFN-gamma expressed in E. coli pIFNTacII is biologically active and routinely recoverable at 10(9) units per liter. When examined microscopically, IPTG induced E. coli harboring either plasmid construction contains prominent cytoplasmic inclusion bodies.     

细胞免疫应答评价方法的研究进展

细胞免疫应答是机体维持稳态的重要组成部分。天然免疫选择性激活适应性免疫后,细胞免疫发挥杀伤清除功能。因此,对细胞免疫应答水平的评估在癌症诊断与治疗、组织器官移植后免疫状态监测、病毒性疾病诊断与预防以及疫苗免疫效果评价等方面意义重大。从最初仅能对体内免疫效果的总体评估,到如今对多种免疫细胞数量和功能的精准检测,细胞免疫应答的评价方法得到了极大发展。然而,细胞免疫应答在机体中涉及多个水平,并且检测方法繁多,难以选择。本文将细胞免疫应答评价方法按照机体、组织器官、免疫细胞和免疫分子4个水平进行系统地整理和比较分析,以期为相关技术的应用提供参考。     

Inability of interferon to protect virus-infected cells against lysis by natural killer (NK) cells correlates with NK cell-mediated antiviral effects in vivo

Murine cytomegalovirus (MCMV) is a natural killer (NK) cell-sensitive virus, whereas lymphocytic choriomeningitis virus (LCMV) is an NK cell-resistant virus. Selective depletion of NK cell activity by injection of mice with anti-asialo GM1 antibody enhanced synthesis of MCMV but not that of LCMV when mice were simultaneously infected with the two viruses. This suggests that the NK cell-mediated antiviral effects may depend on target cell susceptibility to NK cell-mediated lysis rather than the ability of a virus to induce a specialized antiviral NK cell. In support of this concept, activated NK cells isolated from either MCMV- or LCMV-infected mice had similar patterns of killing against all targets tested. Mouse embryonic fibroblasts (MEF) infected with MCMV were less sensitive to lysis by activated NK cells than either uninfected or LCMV-infected MEF. However, when MEF were pretreated with IFN, activated NK cell-mediated lysis against MCMV-infected MEF was undiminished and was much higher (up to fourfold) than that against uninfected MEF, whose sensitivity to lysis was almost totally abolished by IFN pretreatment. LCMV-infected MEF were also protected by IFN against activated NK cell-mediated lysis. During infection, the virus-induced IFN may protect uninfected and LCMV-infected cells from IFN-activated, NK cell-mediated lysis, but MCMV-infected cells may remain sensitive to lysis. This could explain how NK cells play a role in resistance to MCMV but not LCMV.     

Modulation of adaptive immune responses by sphingosine-1-phosphate

Sphingosine-1-phosphate (S1P) has long been recognized as a mediator of a variety of cell functions. A growing body of evidence has accumulated demonstrating its role in cell migration and as a mediator of growth factor-induced events. In recent years, it has become apparent that S1P also mediates many cytokine and chemokine functions. Cells of the immune system function and migrate in response to a complex network of cytokines and chemokines, and the outcome is determined by the interplay of the effects of these molecules on the target cell. S1P may be a bona fide component of these networks and influence the responses of cells to these immune modulators.     

Modulation of immune and inflammatory responses by dietary lipids

PURPOSE OF REVIEW: There continues to be considerable interest in the modulating effect of dietary lipids on immune and inflammatory responses. Although controversy still exists in research in this area, new concepts and approaches have emerged providing useful suggestions. Analysis of the recent findings will help in understanding certain paradoxical findings as well as introducing new strategies to guide future studies. RECENT FINDINGS: The tissue polyunsaturated fatty acid composition was found to be correlated with changes in certain indices of immune function in individuals consuming habitual diets. It seems that individuals or animals with disordered immune systems are more reactive to polyunsaturated fatty acid supplementation, and genetic variation is also a determinant. N-3 polyunsaturated fatty acids were shown to reduce both resistance to bacterial infection and host survival. The studies on other non-classic fatty acids also demonstrated interesting findings. A proposed immuno-enhancing effect of conjugated linoleic acid has not been confirmed by studies and even an adverse effect has been implied. Trans fatty acids have been shown to increase the production of inflammatory cytokines, which may contribute to their pro-atherogenic property. SUMMARY: Current data suggest that the intake of polyunsaturated fatty acids, particularly n-3 polyunsaturated fatty acids, can modulate immune and inflammatory responses, although a discrepancy is still present. Some recent studies have provided useful information explaining possible underlying reasons. Factors such as genetic variation, health status, disease, immune response stage, stimulation type, and possibly age, all contribute to the responsiveness to polyunsaturated fatty acid supplementation in terms of immune function.