T cell help is required to induce idiotypic-anti-idiotypic autoantibody network after immunization with complementary epitope 289-308aa of La/SSB autoantigen in non-autoimmune mice.

Immunotherapies against autoimmune diseases have been of limited success. Preventive vaccines could be developed on the basis to abrogate unwanted immune responses to defined autodeterminants. In this study it is shown that immunization of BALB/c mice with two linear T and B cell epitopes of the human La/SSB autoantigen (spanning the regions 289-308aa and 349-364aa) and their complementary forms specified by the complementary mRNA, results in characteristic B and T cell responses. Mice immunized with the 289-308aa epitope or its complementary peptide elicited specific antibodies against both epitopes. In contrast, mice immunized with the 349-364aa epitope or its complementary peptide mounted antibody titres against the immunizing peptide only. According to these data, the 289-308aa epitope and its complementary form were capable to generate an idiotypic-anti-idiotypic response, which were cross-regulated. Peptide-specific T cell proliferation and cytokine production in vitro revealed the induction of a two-stage T helper response (Th1-->Th2 type) after immunization with either the epitope 289-308 or its complementary peptide. IgG1 was the predominant subclass after immunization with the two forms of epitopes 289-308 and 349-364, while a response of the IgG2b > IgG2a was obtained after the immunization with the complementary form of 349-364 epitope reflecting the TH2/TH1 polarization, respectively. Our data suggest that the complementary peptides of two immunodominant epitopes of human LaSSB can mimic the autoantibodies against these epitopes and establish an active idiotypic-anti-idiotypic network.

Interleukin-1beta and interleukin-1alpha may affect the implantation rate of patients undergoing in vitro fertilization-embryo transfer.

OBJECTIVE: To investigate whether interleukin-1beta (IL-1beta) and interleukin-1alpha (IL-1alpha) affect the implantation rate of patients undergoing IVF-ET. DESIGN: Follicular fluid and serum were obtained on the day of hCG administration, the day of oocyte retrieval, and the day of embryo transfer. SETTING: Cellular immunology laboratory in a research institute, a high technology IVF unit in a medical center, and a university hospital. PATIENT(S): Thirty-three women who were undergoing IVF-ET. MAIN OUTCOME MEASURE(S): IL-1beta and IL-1alpha were measured by specific ELISA and their levels were correlated with the implantation rate. RESULT(S): Classification of IVF-ET patients according to their implantation rate revealed significantly higher amounts of follicular fluid IL-1beta in the implantation versus nonimplantation cycles (68.5+/-24.6 pg/mL versus 20.5+/-13.4 pg/mL); The difference between the level of IL-1alpha in the two groups was not statistically significant(11.6+/-5.1 pg/mL versus 7.3+/-1.9 pg/mL). In parallel, systemic FSH/hMG-dependent IL-1beta and IL-1alpha production was observed in implantation cycles but not in nonimplantation cycles. Statistically significant IL-1beta and IL-1alpha production was observed after administration of hCG. CONCLUSION(S): Gonadotropins used during IVF-ET induce local and systemic production of IL-1beta and IL-1alpha. In addition, the implantation rate for IVF-ET patients who have detectable serum concentrations of IL-1beta and IL-1beta on the day of hCG administration could be higher than the rate for IVF-ET patients who do not have detectable concentrations of these cytokines.

Alteration in peripheral blood mononuclear cell function and serum cytokines in oral lichen planus.

Different activation parameters of peripheral blood mononuclear cells (PBMC) from 31 patients with oral lichen planus (OLP) were examined and compared with 23 healthy donors. Impaired spontaneous (450 +/- 241 vs 1290 +/- 480 cpm) and mitogen-induced (39580 +/- 14470 vs 67000 +/- 11810 cpm) lymphocyte blastogenesis was observed in OLP patients. Furthermore, reduced cytokine production was found after phytohemagglutinin A (PHA) stimulation for all cytokines studied-tumour necrosis factor alpha (TNF alpha, 432.2 +/- 73.4 vs 979.8 +/- 46.3 units/ml), interleukin 2 (IL-2, 156.2 +/- 14.9 vs 572.6 +/- 12.9 pg/ml), interferon gamma (IFN gamma, 48.5 +/- 11.9 vs 82.6 +/- 12.4 pg/ml) and interleukin 6 (IL-6, 253.6 +/- 57.7 vs 1,419.0 +/- 279.6 units/ml)-except for interleukin 1 beta (IL-1 beta) and lymphotoxin (LT). In contrast, unstimulated culture supernatants showed increased TNF alpha (38.2 +/- 13.1 vs 8.0 +/- 0.2 units/ml), LT (10.2 +/- 2.2 units/ml vs < 0.4) and IL-6 (18.5 +/- 5.6 units/ml vs < 0.5) activity. Similarly, elevated concentrations of TNF alpha (19.6 +/- 6.3 units/ml) and IL-6 (22.9 +/- 4.7 units/ml) were detected in the sera of OLP patients. Combination of PHA and phorbol myristate acetate (PMA) could restore OLP proliferative T cell response and cytokine production to the level of healthy donors, whereas exogenous recombinant human IL-2 (rhuIL-2) plus PMA did not seem to be an effective stimulant for OLP T cells. These results indicate an alteration in the immune condition of OLP patients and an impairment in T lymphocyte function.

Anti-inflammatory and immunomodulating effects of the novel agent gamma-(2-aminoethylamino)-2-butyrothienone. 3rd communication: effect on suppressor cells and cytokines production in normal and adjuvant arthritic rats.

The novel agent gamma-(2-aminoethylamino-2-butyrothienone (gamma-ABT) has been found to suppress adjuvant induced disease (AID). To explore the mechanism of action of gamma-ABT in AID, the effects of gamma-ABT on suppressor cells, found in AID, and on interleukin production were evaluated. gamma-ABT did not affect the concavalin A (Con A) responses of AID splenocytes when the suppressor plastic-glass and glass-only-adherent cells were removed. gamma-ABT affected the Con A responses of glass-adherent cells indicating that this agent is selectively affecting the glass-adherent suppressor cells. Interleukins 1 and 2 production is augmented by gamma-ABT in normal and AID rats. Since IL-1 abrogated the suppressive effect of glass-adherent suppressor cells, it is suggested that the inhibition of IL-1 mediated events of suppressor cells may be one of the mechanisms of action of gamma ABT in AID. The relevance of the present results to a latent virus etiology of AID is discussed.

Anti-inflammatory and immunomodulating effects of the novel agent gamma-(2-aminoethylamino)-2-butyrothienone. 2nd communication: inhibitory effect on rat adjuvant induced disease.

The effect of the novel agent gamma-(2-aminoethylamino)-2-butyrothienone (gamma-ABT) on local and systemic changes of rats with adjuvant induced disease (AID) was investigated, gamma-ABT showed potent inhibitory effect on adjuvant primary inflammation and almost totally inhibited the secondary lesions. gamma-ABT improved the changes in lymphoid organ weight except of the thymus. gamma-ABT improved the change in albumin/globulin ratio, which is a parameter of systemic inflammatory reaction but did not improve the body weight gain in AID and normal rats. In addition gamma-ABT did not affect directly T or B lymphocytes as shown by the lymphocyte responses to mitogen (Con A) and a T cell dependent antigen (SRBC) but rather inhibited the suppressor cells found in AID. Although structurally gamma-ABT is completely different from known non-steroidal anti-inflammatories, immunosuppressive drugs behave to a great extent like them.

Interleukins counteract opioid agonists immunosuppression.

A centrally acting novel analgesic, PM, added to murine lymphocyte cultures abrogated the mitogenic response to Con A, as well as, interleukin 2 production, in a dose-dependent manner. Simultaneous presence of interleukin 1 (IL-1) and interleukin 2 (IL-2) into the cultures counteracted, in a dose-dependent manner, as naloxone does, the immunosuppressive action of PM as well as of Pethidine, a known opioid agonist. These results show that IL-1 and IL-2 together exhibit classical agonist-antagonist opioid receptor interactions and support the hypothesis that these lymphokines might play a role as endogenous opioid receptor antagonists against endogenous opioid agonists.

Regulation of isotype immunoglobulin and interleukin production by adjuvants in vitro.

An in vitro system has been developed in which antibodies to fluorescein isothiocyanate (FITC)-labelled human gamma globulin (HGG) or dextran sulfate (DXS), are produced in the presence or absence of different adjuvants. The antibody response of in vitro cultures was measured by assaying the total Ig-secreting cells and FITC-specific plaque-forming cells (PFC). The presence of low levels of antigen and various cytokines were necessary for the production of isotypes other than IgM. Our results indicate that the regulation of isotype switching in vitro is dependent upon the non-specific stimuli-adjuvants, which probably activate different types of cells for cytokine production. The pre-activation of spleen cells, in our system, by antigen, seems to play a decisive role in the subclass of IgG produced. The adjuvant may still be able to influence the precommitted cell isotype to switch to another subclass but only in a "down-stream" direction i.e., IgG3----IgG1----IgG2b----IgG2a.

Regulation of isotype immunoglobulin production by adjuvants in vivo.

Mice were immunized against fluorescein isothiocyanate (FITC)-labelled human gamma globulin (HGG) or dextran sulphate (DXS) in the absence or presence of different adjuvants. The immune response was assayed as the total Ig-secreting cells and FITC-specific plaque-forming cells (PFC) found in various lymphoid organs. The adjuvants influenced the isotype of antibodies produced to the same antigenic determinant. The PFC of different IgG subclasses were favoured by different adjuvants. The IgG3 isotype was produced mainly after immunization with either antigen and lipopolysaccharide (LPS) or Li salt as adjuvant; IgG1 was produced with incomplete Freund's adjuvant (IFA), complete Freund's adjuvant (CFA), alum, poly I:C, Quil A, Be salt, and poly A:U. Some of the above adjuvants (Be salt and poly A:U) favoured the production of IgG2b, and others (CFA, alum, Quil A, and poly I:C) favoured the IgG2a isotype besides the main isotype. Attempts were made to correlate the activation by the various adjuvants of certain TH subtypes with the isotypes produced.