Targeting IL-23 by employing a p40 peptide-based vaccine ameliorates murine allergic skin and airway inflammation.

BACKGROUND: Studies have found that the IL-23/Th17 pathway plays an important role in the pathogenesis of atopic dermatitis (AD) and severe and steroid-resistant asthma. Targeting IL-23/Th17 pathway with monoclonal antibodies (mAb) has been successful in the reduction of skin and airway inflammation in animal models. However, the mAb has a short half-life, requiring repeated administrations. For the long-term suppression of IL-23/Th17 pathway, we have previously developed an IL-23p40 peptide-based virus-like particle vaccine, which induces long-lasting autoantibodies to IL-23. OBJECTIVE: We sought to evaluate the effects of this IL-23p40 peptide-based vaccine on the down-regulation of allergic skin and airway inflammation in mice. METHODS: Mice were subcutaneously injected three times with the IL-23p40 vaccine, or the vaccine carrier protein or saline as controls. Two weeks later, mice were epicutaneously sensitized with ovalbumin four times at a 2-week interval. One week after the final sensitization, mice were nasally administrated with ovalbumin daily for 3 days. One day later, bronchoalveolar lavage fluids (BALF), sera, lung and skin tissues were obtained and analysed. RESULTS: Mice immunized with the vaccine produced high levels of IgG antibodies to IL-23, p40 and IL-12 that in vitro inhibited IL-23-dependent IL-17 production. The numbers of total cells, neutrophils, and eosinophils in BALF were significantly reduced in the vaccine group, compared with controls. The levels of IL-13, IL-5, IL-23 and, IL-17 in BALF and levels of serum ovalbumin-specific IgE, IgG1, and total IgE were also significantly decreased. Histological analysis showed less inflammation of the lung and skin tissues in the vaccine group, compared with controls. CONCLUSION AND CLINICAL RELEVANCE: Administration of an IL-23p40 peptide-based vaccine down-regulates allergic skin and airway inflammation, suggesting that this strategy may be a potential therapeutic approach in the treatment of AD and asthma.

Insidious rifampin-associated renal failure with light-chain proteinuria.

A patient who was receiving rifampin treatment for tuberculosis developed heterogenous light-chain proteinuria and insidious renal failure after a period of fluid restriction. The renal damage was characterized pathologically by an interstitial nephritis with invasive tubular casts and an associated renal vein thrombosis. The possible role of the light-chain proteinuria in the pathogenesis of the renal failure is discussed.

The characterization of a human B cell line utilizable for the assay of B cell growth factors.

An EBV-transformed B cell line JR-2(82) is described which proliferates in response to human B cell growth factor (BCGF) preparations. Although spontaneous replication occurs at high cell densities, at cell densities of 10(3) cells or less, cell death occurs in the absence of BCGF. This response is not dependent upon the presence of low concentrations of supplemental serum in the culture medium. There is no proliferative response to various preparations of interleukin (IL)-1, IL-2 or gamma-interferon (gamma-IFN) up to 1000 U/ml either alone or in combination and specific antisera to IL-2 and gamma-IFN do not interfere with the proliferative response. At 5000 U/ml of gamma-IFN or greater, a partial proliferation of the line is seen to 30% of maximal. The JR-2(82) line and its clones appear to proliferate preferentially in response to low molecular weight (LMW) BCGF, compared to the response seen to high molecular weight (HMW) BCGF from Namalwa cell line supernatants. The line does not produce Ig spontaneously but on incubation with B cell differentiation factor (BCDF) containing supernatants differentiates to an IgG-producing cell line. JR-2(82) or its derivatives may therefore provide a simple reproducible assay for BCGF in the presence of other lymphokines and may be used to study further the interactions of lymphokines in the regulation of the proliferation of EBV-transformed cells.

A comparison of the direct and indirect leukocyte migration tests in man.

In the Direct Leukocyte Migration Test, as few as 10% mononuclear cells are sufficient to produce antigen-specific inhibition of polymorphonuclear leukocyte migration in the presence of 100 mug of excipient-free PPD per ml. A clear distinction between tuberculin-positive and tuberculin-negative donors can be made. The inhibition of migration is blocked by the presence of actinomycin D. In the Indirect Test, significant inhibition of migration of pure polymorphs is produced by supernatants from stimulated tuberculin-sensitive lymphocyte cultures. Inhibition is maximal at 6 h and is of a similar degree to that obtained in the direct test.

Are mitogenic sera necessary for the generation of human plaque-forming cells?

Human plaque-forming cells (PFC) have been quantitated following pokeweed mitogen stimulation by a protein A plaque technique using human erythrocytes. The PFC are generated in cultures supplemented with non-mitogenic human AB serum of foetal bovine serum (FBS) of varying degrees of inherent mitogenicity. In the latter case, no correlation exists between the mitogenicity of the serum supplement used in these cultures and the PFC response obtained. This fact, in addition to the demonstrated usefulness of the AB sera in facilitating the generation of PFC makes it unlikely that serum mitogenicity is a requirement for in vitro human immunoglobulin synthesis and secretion.

Hormones in self tolerance and autoimmunity: a role in the pathogenesis of rheumatoid arthritis?

Recent studies indicate that pituitary hormones play an important role in immunoregulation. The evidence that endocrine abnormalities are associated with, and may contribute to the development of autoimmune disease is reviewed and discussed. Patients suffering from rheumatoid arthritis show a number of endocrine abnormalities that indicate altered pituitary function. The decreased bioactivity of prolactin and possible inadequate glucocorticoid response to inflammation found in patients may have an etiological role in rheumatoid arthritis. The further clarification of the possible role of endocrine factors in the etiology of autoimmune disease is needed urgently.

Cell-mediated allergy to a cerebral aneurysm clip: case report.

BACKGROUND AND IMPORTANCE: Titanium aneurysm clips have superior imaging characteristics and have been supplanting their stainless steel counterparts. We report the case of a 36-year-old woman with a histologically demonstrated cell-mediated reaction to a cobalt alloy aneurysm clip. CLINICAL PRESENTATION: The patient underwent a craniotomy and clip placement for a ruptured cerebral aneurysm. Both the aneurysm clip and the wires used to fix the craniotomy were made of stainless steel (cobalt and nickel) alloys. Postoperatively, the patient was plagued by intense pruritus. INTERVENTION: Skin patch testing revealed a severe reaction to nickel and cobalt. Surgical removal of the aneurysm clip and wires resulted in the disappearance of the patient's symptoms. A histological analysis of the tissue adjacent to the clip and wires was consistent with Type IV delayed type hypersensitivity. CONCLUSION: Stainless steel surgical implants in the head can cause diffuse, severe pruritus in susceptible individuals. Although the role of the aneurysm clip, as opposed to the more superficial wires, in the genesis of the patient's symptoms is not certain, this case demonstrates that a cobalt alloy aneurysm clip can activate T cells and cause an immunological reaction in the central nervous system.

Synovial T lymphocytes in rheumatoid arthritis.

The presence of interleukin-2 (IL-2) like activity has been demonstrated in synovial fluids from rheumatoid arthritis patients. Continuous T cell lines (CTL) generated from such fluids respond to the synovial IL-2, exhibit a predominant suppressor/cytotoxic cell phenotype and release immunoregulatory mediators.

Interaction of a lymphokine with normal human macrophages results in release of a suppressor factor for mitogen-induced immunoglobulin synthesis.

Normal human macrophage/monocyte cultures exposed to a suppressor factor produced by concanavalin A-activated T cells (T-SF), respond by releasing after 72 h a macrophage-derived suppressor factor (M phi-SF). The M phi-SF inhibits pokeweed mitogen-induced Ig synthesis but not T- or B-cell proliferation. Cycloheximide treatment of the macrophages does not interfere with generation of the M phi-SF, suggesting that de novo synthesis is not required. The factor is not preformed, for virgin macrophages do not contain M phi-SF, but it appears in macrophage cell lysates after exposure to T-SF. The production of the M phi-SF is inhibited by the presence of 2-mercaptoethanol. Both T-SF and M phi-SF are L-rhamnose inhibitable, and the M phi-SF appears to be released as a high molecular weight complex which is dissociable into a low molecular weight form of a size similar to the T-SF, i.e. approximately 20,000. The T-SF induced M phi-SF has some similarities with soluble immune response suppressor (SIRS) but differs from this factor in its lack of effect upon lymphocyte proliferation failure to induce conversion of T-SF to M phi-SF by treatment with H2O2.

Interleukin-2 abnormalities in systemic lupus erythematosus and rheumatoid arthritis. A role for overproduction of interleukin-2 in human autoimmunity?

Interleukin-2 (IL-2) production in systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA) was assessed at varying cell densities from 3 X 10(3)-1 X 10(5) cells/culture in response to mitogen stimulation, using a cellular interleukin assay in which IL-2 responsive CTLL-2 indicator cells were added directly to stimulated peripheral blood lymphocyte cultures. It was found that in both RA and SLE there was a significant overproduction of IL-2 demonstrable at low cell densities. At high cell densities, IL-2 production was decreased in SLE but remained enhanced in RA. The former abnormality appears to be related to disease activity and was not reversed by irradiation, suggesting that it is not the result of the actions of a radiation sensitive suppressor cell population. The possibility is discussed that a local hyper-production of IL-2 could induce disturbances of the immune response, resulting in the breaking of normal tolerance.

B cell differentiation factor production in systemic lupus erythematosus.

Spontaneous and mitogen stimulated B cell differentiation factor (BCDF) production by peripheral blood lymphocytes (PBL) from patients with systemic lupus erythematosus (SLE) was compared with that of normal control subjects. BCDF production was measured by a cellular interleukin assay in which a BCDF responsive cell line was cocultured with increasing numbers of PBL from the donor populations. It was found that no significant differences occurred in BCDF production by SLE and normal PBL under the conditions used. Indeed there was a trend toward decreased BCDF release from SLE PBL. This would support the hypothesis that human SLE is the result of enhanced B cell responsiveness to helper cell factors rather than due to increased release of such factors.

Repeat skin testing for aeroallergen immediate hypersensitivity in an atopic patient population.

A retrospective study of repeat intradermal skin testing with aeroallergens carried out on atopic subjects with allergic rhinitis/conjunctivitis and/or asthma, after a mean interval of 3.5 years, showed that an increase in the number of allergens reacted to by an individual patient correlated with age or with worsening of symptoms.

The production of immunoregulatory factors by a human macrophage-like cell line. III. The relationship between the lymphoid-specific inhibitor of DNA synthesis and gamma-interferon.

Mitogenic stimulation of sublines of the human histiocytic lymphoma, U937, results in the production of an inhibitor of lymphocyte proliferation (IDS-U937). The relationship of IDS-U937 to alpha- and gamma-interferon was examined by several techniques: (1) the antiviral activity of the IDS preparations was determined, (2) the effects of antibodies to interferon on IDS activities, and (3) the comparison of the antiproliferative activities of gamma-IFN and IDS. The relationship of IDS to other macrophage derived inhibitors is also discussed with respect to target cell specificities and modes of action.

Lymphocyte-mediated cytotoxicity in isoniazid-associated hepatitis.

The release of lymphotoxin (LT) from peripheral blood lymphocytes of patients with isoniazid (INH)-induced hepatitis was studied, using L929 fibroblast target cells, as was the cytotoxic effect of these lymphocytes on murine hepatoma cells (L1469) and L929 fibroblasts, using a 3H-proline cytotoxicity assay. Evidence for LT release was found in five out of six patients, following stimulation of the peripheral blood lymphocytes with INH or isonicotinic acid (INA) conjugated to human serum albumin. In the direct cytotoxicity assay, cytotoxic effects on the hepatoma cells were enhanced by preincubation of the target cells with INH in five out of six patients tested. Although specificity with regard to the drug was demonstrable, tissue specificity was less certain in that enhanced killing of the fibroblast cell line was also found to occur following preincubation of the L929 cells with INH.

Lymphocyte transformation studies in drug hypersensitivity.

In a group of patients with clinically diagnosed drug hypersensitivity the in vitro lymphocyte response to the suspected drug was assessed by the lymphocyte transformation test. The test gave positive results in all 15 patients with penicillin-induced immediate or accelerated allergic reactions and positive immediate skin-test reactivity to the major or the minor antigenic determinant of penicillin, or both, but in only 3 of the 12 patients with delayed-onset maculopapular rashes induced by penicillin, despite positive immediate reactivity to the skin-test reagents.Lymphocyte stimulation greater than five times the control level was demonstrated for five patients with penicillin-induced erythroderma, Stevens-Johnson syndrome or a serum-sickness-like illness, or with methicillin-induced interstitial nephritis, all of whom had negative reactions to the appropriate skin-test reagents. A low level of stimulation was seen in eight other skin-test-negative patients with possible allergic reactions induced by penicillins. However, in all subjects tested the stimulation was significantly greater than the mean for control subjects.For 9 of 11 patients with isoniazid-induced hepatitis or maculopapular rashes, but for only 8 of 31 patients with eruptions induced by a variety of drugs other than penicillins and isoniazid, significant stimulation occurred in the lymphocyte transformation test.It is concluded that the lymphocyte transformation test is useful in the detection of hypersensitivity to the penicillins (although in IgE-mediated reactions skin testing is clearly preferable) and isoniazid but is of limited value in the demonstration of hypersensitivity to other drugs.

Polymer modified electrodes for the reversible oxidation-reduction of NAD+/NADH for use within amperometric biosensors.

Electrochemical enzyme based biosensors continue to attract much attention due to the simplified analyses they promise, in comparison to many more complicated analytical procedures. The largest group of enzymes, are those that rely NAD(P)+/NAD(P)H as cofactors. One of the problems associated with the use NAD(P)+/NAD(P)H redox couples, however, within electrochemical biosensors is their ability to passivate noble metal electrodes. There have been many attempts to overcome this problem, such as the use of carbon paste electrodes [1], although approaches such as these are both cumbersome and typically irreproducible. In recent years conducting polymers such as poly(3-methylthiophene) [2] have also been used to modify noble metal electrode surfaces to lower the overpotentials required, while poly(phenol red) has been incorporated into films to offer selectivity against interferents [3,4]. In this study platinum electrodes have been electrochemically coated with mixed polymer films of poly(phenol red) together with poly(3-methylthiophene), poly(aniline) or poly(o-phenylenediamine) to offer enhanced performance. Cyclic voltammetry has been used to compare the electrochemical behaviour of the NAD+/NADH redox couple over repeated potential cycles, with the most favourable results being seen with the poly(aniline)/poly(phenol red) and poly(o-phenylenediamine)/poly(phenol red) coated electrodes.

Normal mitogen-induced suppression of the interleukin-6 (IL-6) response and its deficiency in systemic lupus erythematosus.

A low-frequency suppressor-cell population in normal peripheral blood inhibits the B-cell CESS response to IL-6, following pokeweed mitogen stimulation. The suppression of IL-6 responsiveness is (i) radiation sensitive, (ii) directed against CESS targets and not mediated by inhibition of IL-6 production, and (iii) associated with nonspecific cytotoxic activity against CESS targets. The generation of these cytolytic cells is also radiation sensitive. A correlation was found between PWM-induced cytotoxicity against CESS and the suppression of IL-6-dependent IgG production. But cytotoxicity toward CESS targets is not responsible for this suppression because (i) IL-2 induces equivalent or greater nonspecific cytotoxicity against CESS in the total absence of suppression of CESS-derived IgG production and (ii) suppression is also induced by mitogen-activated PBL separated from CESS targets by a cell-impermeable membrane. This suppression was not mediated by TNF alpha/beta or IFN-gamma. In systemic lupus erythematosus, suppression of IL-6-dependent IgG production is impaired in patients with active disease (29.2 +/- 13.7%) compared to patients with inactive disease (70 +/- 19.5%) or normal controls (82.8 +/- 9.2%). There is also a defect in mitogen-induced nonspecific cytotoxicity in active SLE (specific lysis 15.1 +/- 3.5%, compared to 34 +/- 4% in normals). Pokeweed mitogen-activated PBL can therefore normally induce suppression of B-cell IL-6 responses and this response is deficient in lupus.

Lymphokine production in rheumatoid arthritis and systemic lupus erythematosus.

The production of interferon (IFN) by peripheral blood lymphocytes from patients with rheumatoid arthritis (RA) and of IFN and interleukin 2 (IL-2) in systemic lupus erythematosus (SLE) was compared with that of healthy controls. Patients with SLE showed a significant reduction in IL-2 production compared to controls if the PBL were irradiated before mitogen stimulation. No patient with RA or SLE studied had impaired IFN production regardless of disease activity and the IFN produced was always IFN-gamma in type. We conclude that there is an abnormality in IL-2 production in SLE but there is no abnormality in IFN-gamma production in either RA or SLE.

Severe delayed-onset hypersensitivity reactions to amoxicillin in children.

Amoxicillin, a semisynthetic aminopenicillin, has achieved widespread use in recent years for the treatment of respiratory tract and otic infections. Serious reactions have been relatively infrequent. From July 1986 to June 1987, 11 children aged 6 months to 10 years presented with delayed-onset hypersensitivity reactions. In 10 the symptoms were consistent with a serum-sickness-like illness, including urticaria, angioedema, arthritis and arthralgia. Radioallergosorbent testing for IgE antibodies to penicillin yielded negative results, and lymphocyte transformation testing gave a positive result in only one patient. Because of the negative immunologic test results and the occurrence of reactions only in children, who had received an amoxicillin solution, the reactions may have been caused by the excipient.