Altered intracellular expression of the chemokines MIP-1alpha, MIP-1beta and IL-8 by peripheral blood CD4+ and CD8+ T cells in mild allergic asthma.

BACKGROUND: The ability of chemokines to regulate Th1 and Th2 responses suggests a role in the pathogenesis of atopic disorders such as allergic asthma where Th2 response dominance has been observed. Although the impact of allergic asthma on local chemokine production in the lung has been the subject of investigation, little is know about the influence of disease progression on peripheral chemokine production. We now report use of whole blood culture and flow cytometry to assess the influence of mild allergic asthma on peripheral T-cell chemokine expression. METHODS: Study participants included patients with mild allergic asthma (n = 7) and nonasthmatic controls (n = 7). Following in vitro stimulation of peripheral venous blood with phorbol 12-myristate acetate (PMA) and ionomycin, flow cytometry was used to estimate the percentage of CD4+ and CD8+ T cells producing a number of chemokines, including macrophage inflammatory proteins MIP-1alpha and MIP-1beta, RANTES (regulated on activation, T-cell expressed and secreted), monocytic chemotactic protein-1 (MCP)-1, and interleukin (IL)-8, or the cytokines interferon (IFN)-gamma and IL-4. Serum levels of MIP-1alpha, MIP-1beta, RANTES, MCP-1, IL-8, IFN-gamma and IL-4 were also assessed by quantitative ELISA. RESULTS: Intracellular expression of MIP-1beta by CD4+ and CD8+ T cells from allergic asthmatics was significantly reduced in comparison to that observed for nonasthmatics (median = 2.29% (1.75-3.50) vs 4.57% (3.38-6.64), P = 0.05; 14.20% (13.18-17.88) vs 44.10% (30.38-48.70), P = 0.01). Similarly, intracellular expression of MIP-1alpha by CD8+ T cells from allergic asthmatics was also significantly lower (3.67% (1.17-5.42) vs 17.10% (4.97-20.43), P = 0.05). Conversely, IL-8 expression by both CD4+ and CD8+ T cells from allergic asthmatics demonstrated significant enhancement (9.93% (7.77-11.28) vs 4.14% (3.61-7.11), P = 0.05; 8.40% (6.97-10.04) vs 4.98% (3.37-6.08), P = 0.05). Examination of intracellular IFN-gamma and IL-4 revealed no significant difference in the expression of either cytokine by CD4+ T-cells from allergic asthmatics and nonasthmatics. In contrast, expression of IFN-gamma was significantly reduced in CD8+ T-cells from allergic asthmatics (24.60% (21.08-32.50) vs 48.40% (41.50-55.28), P = 0.01). CONCLUSIONS: The occurrence in mild allergic asthma of peripheral T-cell chemokine expression suggestive of a diminished Th1 response, coinciding with marginal change in cytokine profiles indicative of a Th2 response bias, confirms the importance of chemokine involvement in the etiology of allergic asthma. The ability to use whole blood culture to estimate chemokine expression in T cell subsets may ultimately provide a practical means to evaluate disease status and to monitor early intervention therapies which target chemokines.

Long-term evaluation of T-cell subset changes after effective combination antiretroviral therapy during asymptomatic HIV-infection.

Demonstration of long-lived HIV-reservoirs resistant to the effects of combination antiretroviral therapy raises concern over the ability of treatment to maintain long-term beneficial alterations in T-cell subset composition. To address this issue, we have examined the effect of antiretroviral therapy on T-cell subset change during early HIV-infection in a 2-year prospective open-label trial composed of treatment-naive asymptomatic HIV-infected patients with CD4+ T-cell counts > or =400 cells/microl. Therapy consisted of double (zidovudine and lamivudine) or triple (zidovudine, lamivudine, and ritonavir) combination antiretroviral therapy. Retrospective analysis based on magnitude of viral suppression was used to characterize responder and nonresponder groups. Among responders, long-term antiretroviral therapy maintained a significant increase in numbers of total CD4+, naive CD4+/CD45RA+, and memory CD4+/CD45RO+ T cells. A concomitant significant decrease in numbers of memory CD8+/CD45RO+ and both activated CD8+/HLA-DR+ and CD8+/CD38+ T cells was also maintained. In contrast, long-term antiretroviral therapy among nonresponders led only to a significant increase in the numbers of CD4+ T cells and a significant reduction in numbers of activated CD8+/HLA-DR+ T cells. The long-term ability of antiretroviral therapy during early asymptomatic HIV-infection to maintain reversal of disease-induced T-cell activation and maturation abnormalities continues to support the concept that immunologic advantage is gained by commencing early aggressive antiretroviral therapy. Nevertheless, continued management of T-cell subset recovery is significantly more effective in the presence of completely suppressed viral replication.

Change in circulating levels of the chemokines macrophage inflammatory proteins 1 alpha and 11 beta, RANTES, monocyte chemotactic protein-1 and interleukin-16 following treatment of severely immunodeficient HIV-infected individuals with indinavir.

OBJECTIVE: To evaluate the in vivo relationship between HIV replication and circulating levels of the chemokines macrophage inflammatory protein (MIP)-1 alpha, MIP-1 beta, RANTES (acronym for Regulated upon Activation, Normal T-cell Expressed and presumably Secreted), interleukin (IL)-16 and monocyte chemotactic protein (MCP)-1, which have recently been characterized as factors capable of regulating in vitro HIV replication. DESIGN AND METHODS: We have compared changes in plasma HIV-RNA levels and circulating levels of MIP-1 alpha, MIP-1 beta, RANTES, IL-16 and MCP-1 in 20 severely immunodeficient HIV-infected individuals (CD4+ T cells = 14 +/- 3 cells x 10(6)/l; plasma HIV RNA = 4.45 +/- 0.27 log 10 copies/ml) undergoing treatment with the HIV protease inhibitor indinavir that added to pre-existing nucleoside-based therapy. At weeks 0, 2, 6 and 12, viral load was quantified using a commercial reverse-transcription polymerase chain reaction assay, peripheral blood T-cell subpopulations assessed by flow cytometry, and chemokine levels quantified using commercial sandwich enzyme-linked immunosorbent assay kits. RESULTS: Following initiation of indinavir-based therapy, significant decreases in plasma HIV-RNA levels (change = 2.0 +/- 0.75 log 10 copies/ml) were observed in conjunction with significant increases in absolute CD4+ (change = 83 +/- 19 cells x 10(6)/l) and CD8+ (change = 293 +/- 96 cells x 10(6)/l) T-cell numbers. Concomitantly, significant increases in MIP-1 alpha (19% increase), MIP-1 beta (14% increase), RANTES (15% increase) and IL-16 (1213% increase) levels occurred. In contrast, MCP-1 levels decreased significantly (47% decrease). CONCLUSION: The in vivo demonstration of an association between diminishing plasma HIV-RNA levels and the emergence of a circulating chemokine profile capable of inhibiting HIV replication corroborates recent in vitro observations and provides evidence for the restoration of chemokine capacity by HIV protease inhibitor-based therapy.

Interleukin-8 released into the cerebrospinal fluid after brain injury is associated with blood-brain barrier dysfunction and nerve growth factor production.

Interleukin (IL) 8 was measured in CSF of 14 patients with severe traumatic brain injury. IL-8 levels were significantly higher in CSF (up to 8,000 pg/ml) than serum (up to 2,400 pg/ml) (p < 0.05), suggesting intrathecal production. Maximal IL-8 values in CSF correlated with a severe dysfunction of the blood-brain barrier. Nerve growth factor (NGF) was detected in CSF of 7 of 14 patients (range of maximal NGF: 62-12,130 pg/ml). IL-8 concentrations were significantly higher in these patients than in those without NGF (p < 0.01). CSF containing high IL-8 (3,800-7,900 pg/ml) induced greater NGF production in cultured astrocytes (202-434 pg/ml) than samples with low IL-8 (600-1,000 pg/ml), which showed a smaller NGF increase (0-165 pg/ml). Anti-IL-8 antibodies strongly reduced (52-100%) the release of NGF in the group of high IL-8, whereas in the group with low IL-8, this effect was lower (0-52%). The inability of anti-IL-8 antibodies to inhibit the synthesis of NGF completely may depend on cytokines like tumor necrosis factor alpha and IL-6 found in these CSF samples, which may act in association with IL-8. Thus, IL-8 may represent a pivotal cytokine in the pathology of brain injury.

[Cost-benefit relationship of serologic studies in autoimmune diseases]. / Kosten-Nutzen-Verhältnis der serologischen Untersuchungen bei autoimmunen Erkrankungen.

Autoimmune disorders are mainly clinically defined disease entities. The diagnosis of many of the systemic autoimmunopathies is based on characteristic combinations of symptoms, some of them overlapping various kinds and being of unspecific nature. Therefore, laboratory parameters such as autoantibodies, components of the complement system, some cytokines and their receptors, etc., are used for the diagnosis, although many of them are of limited specificity and sensitivity. The most important immunoparameters being of value for diagnosis and disease monitoring are discussed, and a stepwise procedure is proposed in order to reduce the costs. Basis are the screening tests for antinuclear antibodies (ANA) and for antibodies against cytoplasmatic components of granulocytes (ANCA). Depending on more specific questions, other tests and test combinations are then applied in second and third priorities.

[Antineutrophil cytoplasma antibodies (ANCA): clinical indications and experience with these new autoantibodies]. / Anti-Neutrophilen-zytoplasmatische Antikörper (ANCA): Indikationsbereich und Erfahrungen mit diesem neuen Autoantikörper.

Antineutrophil cytoplasmatic autoantibodies (ANCA) have been tested in this laboratory for more than two years with a 3-fold increase in tests and positive results. The initial association with Wegener's granulomatosis has since been extended to microscopic polyarteritis and rapidly progressive glomerulonephritis. We review the published data. ANCA are not simply another laboratory test but have become an important tool for diagnosis, treatment monitoring and prevention of relapses in many vasculitis syndromes including some forms of rapidly progressive glomerulonephritis. The ANCA-associated antigens have been identified as a serin-proteinase and myeloperoxidase. This provides insights into the pathogenesis of the ANCA-related diseases.

Measurement of proteins with the Behring Nephelometer. A multicentre evaluation.

The selective multi-protein analyser Behring Nephelometer was examined according to the ECCLS guidelines in a multicentre evaluation involving five laboratories. IgG, IgA, IgM, C-reactive protein, C3c, C4, apolipoprotein A-I and B were measured in serum, and albumin and IgG were measured in cerebrospinal fluid. All values obtained were included in the evaluation without correcting for outliers. The trial, which lasted three months and involved over 20,000 analyses, basically yielded the following results: 1. The precision was generally better than that of comparative procedures. For the majority of methods, the between-day coefficients of variation were below 4.5%. The highest coefficient of variation was 7.2% (for C-reactive protein) and the lowest 1.35% (for C3c). 2. The fraction of assigned values of control materials varied between 0.95 and 1.08. 3. Good agreement was found with results from the comparison procedures: Beckman ICS, Behring Laser Nephelometer, Hyland Laser Nephelometer. 4. No carry-over effects were observed. 5. No interferences were observed for IgG, IgA and IgM determinations using hyperbilirubinaemic or haemolytic samples. In contrast lipaemic samples and some with monoclonal immunoglobulin M showed an influence on the immunoephelometric reaction. 6. Because of the large measuring range, it is necessary to repeat analyses only in extremely rare cases. 7. During the entire evaluation period no instrument malfunctions or interruptions occurred. As a result of its reliability, the Behring Nephelometer is well suited for routine operation and emergency analyses in medium and large-sized laboratories.