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Multiparameter Analysis of Clastogenic Factors, Pro-oxidant Cytokines, and Inflammatory Markers in HIV-1-Infected Patients with Asymptomatic Disease, Opportunistic Infections, and Malignancies

Molecular Medicine volume 4, pages 333–343 (1998)Cite this article

Abstract

HIV-1-infected patients are in chronic oxidative stress and clastogenic factors (CFs) are present in their plasma. CFs from patients with HIV are formed via superoxide anion radical and stimulate further superoxide production. The pathophysiologic significance and the exact composition of the circulating clastogenic material in patients with HIV is unknown. Cytokines, such as tumor necrosis factor-alpha (TNF-α), are increased in the plasma of patients with HIV and TNF-α shows clastogenic activity in vitro. The aim of this clinical study was to compare levels of CF in HIV-1-positive patients with asymptomatic disease, opportunistic infections, and malignancies with those in HIV-1-negative control groups and to correlate CF activity with CD4+ T cell numbers, the cytokines (TNF-α, interleukin-2 [IL-2], IL-6), and the inflammatory markers (C-reactive protein [CRP], neopterin, granulocyte elastase). CFs were significantly increased in all HIV-1 -positive patients and in HIV-1 -negative patients with malignant tumors. HIV-1-positive patients with Kaposi’s sarcoma showed the highest CF activity in their plasma (p < 0.08). CFs appear very early in HIV infection, and they correlate negatively with CD4+ T cells, which are an indicator of disease activity. The presence of CF in the plasma of HIV-infected patients is not a general response to a viral infection because these factors are not increased in HIV-1-negative patients with viral infection (zoster). CFs are not specific for the HIV-1 infection; they also occur in HIV-1-negative patients with malignant tumors. There was a tendency towards a positive correlation (p < 0.14) between CF and TNF-α, but there was no positive correlation of CF with IL-2, IL-6, CRP, elastase, and neopterin levels. This indicates that TNF-α may be among the components of CF in HIV-1-infected patients. In addition, other unidentified components may contribute to the clastogenic activity of the plasma or the composition of CF may vary from patient to patient. Further clinical studies with larger sample populations are necessary to analyze the composition of CF in HIV-1 -positive patients.

Introduction

Clastogenic factors (CFs) are chromosome-damaging agents with low molecular weight (<30,000 daltons) that cause chromosome aberrations, sister chromatid exchanges, DNA strand breakage, and gene mutation. For more than 30 years they have been known to be an indirect effect of ionizing radiation (1,2). Because of their persistence in the blood of irradiated persons many years after exposure (3), CF have been considered risk factors for late effects of ionizing irradiation such as cancer and leukemia (4). The presence of CF in plasma has also been described in patients with congenital breakage syndromes such as ataxia teleangiectasia, Fanconis anemia, and Bloom’s syndrome, which are all associated with a high incidence of malignancies (5). The superoxide anion radical is implicated in clastogenic factor formation and action (6). This has been demonstrated particularly for CF derived from the plasma of patients with human immunodeficiency virus (HIV) (7). Recently, the presence of CF was reported in patients infected with the HIV-1 virus (8).

Experimental evidence indicates that HIV-1 -infected patients are in oxidative imbalance, which starts early in the course of the disease. Cytokines, such as TNF-α (9,10) and IL-6 (11), and acute phase proteins, such as CRP (12) and neopterin (13), are increased in HIV-1-infected patients, while IL-2 (14) is decreased. These products may stimulate the cellular production of reactive oxygen species. Low levels of serum and tissue antioxidants and elevated concentrations of peroxidation products have been reported in HIV-infected patients (15–21). HIV induces immunosuppression that renders the body highly susceptible to opportunistic infections and malignancies. Oxidative stress may contribute to the immunosuppression, carcinogenesis, and stimulation of viral replication in HIV-infected patients (22–25).

The objective of this study was to compare levels of CF in the plasma of HIV-1-positive patients with asymptomatic disease, opportunistic infection (zoster), and malignancies (T cell lymphomas, Kaposi’s sarcoma) with those in HIV-1-negative control groups. We investigated the correlation of CF values with the CD4+ T cells to learn about its indicator function for disease progression. In addition, the plasma levels of the cytokines IL-2, IL-6, and TNF-α and the acute-phase proteins CRP, neopterin, and granulocyte elastase were correlated with CF activity to find out about the factors that may contribute to CF formation.

Materials and Methods

Patients

We examined 30 HIV-1-negative and 24 HIV-1-positive out- and inpatients from the Departments of Dermatology and Internal Medicine, J. W. Goethe University Medical School, Frankfurt, Germany. The HIV-1-negative patients were subdivided into the following groups: HIV-1-negative healthy controls (n = 10, mean age 30 years; range = 23–69 years; no antibiotic or antiviral drugs); HIV-1-negative patients with zoster infection (n = 10, mean age 57 years, range = 36–82 years; no antibiotic or antiviral drugs); and HIV-1-negative patients with malignant tumors (T cell lymphomas, sarcomas), (n = 10, mean age 56 years, range = 21–88 years; no antibiotic or antiviral drugs). The HIV-1-positive patients were stratified into the following groups: HIV-1-positive patients with asymptomatic disease (CD4+ T cell numbers > 500/ml) (n = 10, mean age 36 years, range = 24–50 years); HIV-1-positive patients with zoster infection (n = 4, mean age 53 years, range = 43–67 years); and HIV-1-positive patients with malignant tumors (Kaposi’s sarcoma n = 7, T cell lymphomas n = 3), (n = 10, mean age 44 years, range = 33–63 years). One HIV-1-positive patient with zoster also had Kaposi’s sarcoma. Most of the HIV-1 -positive patients received antibiotic or antiretroviral therapy. The HIV-1-negative patients with malignancies did not receive radiation therapy prior to collection of plasma. Patients and healthy subjects gave their informed consent to participate in this study, according to the Medical School’s Ethics Committee regulations.

Sample Preparation

For evaluation of the clastogenic effects exerted by the patient’s plasma, the blood was centrifuged. The plasma was ultrafiltered through an Amicon filter, which had a cutoff at 30,000 daltons (Amicon YM 30, Danvers, MA). This procedure removes all high-molecular-weight plasma components that might disturb culture growth because of blood group incompatibilities. In addition, it eliminates cells that might still be present in the plasma after centrifugation. The ultrafiltrates were then stored at − 80°C and delivered frozen to the Institut Biomedical des Cordeliers in Paris, where they were tested for their clastogenic activity in a cytogenetic test system. The preparation and analysis were carried out according to the standard procedure reported in detail elsewhere (26).

Clastogenic Factor Test

Aliquots of the ultrafiltrates (0.25 ml) were added to the test cultures set up with 0.5 ml whole blood from healthy blood donors, 5 ml tissue culture medium (TCM 199 from Flow Laboratories, Paris) and 1 ml fetal calf serum (FCS) (Gibco, Paris). In some cases, 0.25 ml of the patients’ ultrafiltrate caused significant cytotoxicity in the donor lymphocytes. Therefore, the assay was repeated with 0.1 ml ultrafiltrate and the highest rates of chromosome aberrations were taken for the calculation of CF. In controls (healthy blood donors), 0.25 ml or 0.1 ml of plasma ultrafiltrates caused the same rate of chromosome aberrations. Lymphocyte proliferation was stimulated by addition of phytohemagglutinin (Wellcome Diagnostics, Dartford, U.K.). After 72 hr of incubation at 37°C, microscopic slides were prepared for chromosomal analysis according to standard procedures. Cultures of normal donor cells showed a small number of chromosomal aberrations, or so-called spontaneous aberrations. This background level of aberrations in the simultaneously untreated blood cultures was substracted from the aberration rate in the ultrafiltrate-treated cultures of the same blood donors. The difference between the two values is given as the level of CF (27). The plasma ultrafiltrates from about 100 healthy blood donors from the Centre de Transfusion in Paris showed a maximum of 6 CFs in 5% of the donors (historical control). A higher rate of CF was never observed, therefore CF > 8 is considered abnormal.

Inflammatory Markers, Cytokines, and CD4+ T Cell Analysis in Plasma

CRP (28) (Behring-Turbi-Time® test, Behring, Marburg, Germany), granulocyte elastase (29) (Immunoassay Ecoline® Merck, Darmstadt, Germany), neopterin (30) (Neopterin Immunoassay Merck, Darmstadt, Germany), TNF-α (Predicta®, Genzyme Diagnostics Inc, Cambridge, MA), IL-2 (Interleukin-2 Immulite®, DPC Biermann, Bad Nauheim, Germany) and IL-6 (Interleukin-6 Immulite®, DPC Biermann, Bad Nauheim, Germany) were determined by commercially available test systems (31–33). CD4+ T cells were analyzed in a Coulter counter for routine hematological analysis.

Statistics

The experimental data points showed a non-Gaussian distribution, therefore nonparametric tests were selected for statistical analysis. The unpaired Kruskal-Wallis test was used for comparison of multiple independent data points, and the Wilcoxon-Mann-Whitney test was applied for comparison of two independent data points. The Spearman rank test was applied as a non-parametric test for correlation analysis. A p < 0.05 was considered significant.

Results

Clastogenic Factors

The plasma ultrafiltrates from HIV-1-positive patients with asymptomatic disease (p < 0.002), with zoster infection (p < 0.01), and with malignancies (p < 0.00008) induced a significant increase in chromosomal breakage in the cytogenetic test system compared with HIV-1 -negative healthy controls. HIV-1-negative patients with malignant tumors also showed a significantly increased CF (p < 0.008). No increase in chromosome breaks was observed in test cultures exposed to plasma ultrafiltrates from HIV-1 -negative patients with zoster infection compared with HIV-1-negative healthy controls (Table 1). HIV-1-positive patients with Kaposi’s sarcoma tended to show the highest CF activity in plasma (16 ± 4.0), compared with all HIV-1-positive patients without Kaposi’s sarcoma (10 ± 1.2; p < 0.08) (Table 1). There was no significant difference in CF in the plasma of smokers versus non-smokers in healthy controls and patients. Furthermore, alcohol intake did not influence the CF in our patients and controls (data not shown).

Table 1 CFs, cytokines, and inflammatory markers in HIV-1-positive patients and control groups
Full size table

Cytokines

The plasma levels of the cytokines TNF-α, IL-2, and IL-6 were generally within the normal range in all patients studied. However, TNF-α concentration tended to be elevated in HIV-1-negative patients with malignancies (p < 0.1) and in HIV-1-positive patients with malignancies (p < 0.09), in comparison with HIV-1-negative controls. HIV-1 -positive patients with malignancies showed the highest TNF-α values (Table 1).

IL-6 was significantly increased in HIV-1-negative patients with malignancies (p < 0.04) as well as in HIV-1-positive patients with asymptomatic disease (p < 0.04) and with zoster (p < 0.04), compared with HIV-1-negative healthy controls. IL-6 was significantly decreased in HIV-1 -positive patients with malignancies in comparison to HIV-1-positive patients with asymptomatic disease (p < 0.0003) (Table 1). IL-2 levels were inclined to be more increased in HIV-1-positive patients with malignancies (p < 0.1) (Table 1) than in HIV-1-negative controls.

Inflammatory Markers

Neopterin levels were significantly increased in HIV-1-negative patients with zoster (p < 0.02) or malignancies (p < 0.03), and in HIV-1-positive patients with zoster (p < 0.01) or malignancies (p < 0.03). Neopterin was not elevated in HIV-1-positive patients with asymptomatic disease (Table 1). Granulocyte elastase and CRP were inclined to be increased in HIV-positive patients with zoster (p < 0.1).

Correlation Analysis

CF tended to correlate negatively with CD4+ T cell numbers (r = −0.37, p < 0.1). This negative correlation reached statistical significance (r = −0.44, p < 0.05) when 0.1 ml plasma ultrafiltrate was used for the cytogentic assay (see Materials and Methods). CF correlated negatively with IL-6 (r = −0.43, p < 0.05) and showed a tendency towards a positive correlation with TNF-α (r = 0.31, p < 0.14) (Table 2). CD4+ T cells correlated positively with IL-6 (r = 0.57, p < 0.01), and CD4+ T cells showed a tendency towards a negative correlation with neopterin (r = −0.37, p < 0.1) (Table 2). TNF-α correlated positively with CRP (r = 0.5, p < 0.02), elastase (r = 0.47, p < 0.05) and neopterin (r = 0.44, p < 0.05) (Table 2). CRP correlated positively with elastase (r = 0.92, p < 0.002) and neopterin (r = 0.79, p < 0.002). Elastase correlated positively with neopterin (r = 0.72, p < 0.002) (Table 2).

Table 2 Spearman rank correlation analysis of CF, CD4 + T cells, cytokines, and inflammatory markers in HIV-1-infected patients
Full size table

Discussion

Clastogenic Factors

CFs are frequently but not exclusively found in several clinical conditions associated with a high incidence of malignancies. CF-induced clastogenesis is comparable to chemical clastogenesis, the only difference being that these clastogens are of endogenous origin. The endogenous clastogens induce chromosome damage not only in the cells of the patient (donor) but also in test cultures set up with blood of healthy persons. The latter is used as an assay for the detection of these clastogens. CFs are not single factors but mixtures of chromosome-damaging pro-oxidant substances. Biochemical analysis of the clastogenic plasma identified various chromosome-damaging agents, e.g., lipid peroxidation products such as 4-hydroxynonenal (34) and inosine triphosphate (35) and a monocyte-derived component (36), which was later identified as TNF-α (6).

The presence of CF in the plasma of HIV-1-positive patients very early in the course of the disease (HIV patients with asymptomatic disease; Table 1) contributes to a shift in the pro-antiox-idant balance toward the pro-oxidant side and may be of pathophysiological significance. Patients will be exposed to the clastogenic effects as long as the vicious cycle of CF and superoxide anion radical production is not interrupted (37). Pro-oxidant states can modulate the expression of genes related to cell growth and differentiation (38), and extracellular superoxide anion generation has been shown to activate proto-oncogenes and induce mutations at tumor suppressor loci via strand breakage and DNA sequence changes (39,40). Whether the clastogenic effects continuously produced by circulating CF in HIV-1-positive patients represents a risk factor for malignancies frequently observed in these patients deserves further study and follow-up. Recently, oxidative stress was suggested to be a viable clinical factor in AIDS Kaposi’s sarcoma development (41). The finding that HIV-1-positive patients with Kaposi’s sarcoma showed the highest CF values of all HIV patients (although statistically not significant, p < 0.08) warrants further investigation. The presence of CF in the plasma of HIV-1 -positive patients is not a general response to a viral infection because CFs were not increased in HIV-1-negative patients with zoster virus infection (Table 1). CF formation is not specific for HIV-1 infection because CF levels were also significantly elevated in HIV-1-negative patients with malignancies (Table 1). The negative correlation of CF with CD4+ T cells numbers (r = −0.37, p < 0.1), which reached statistical significance (r = −0.44, p < 0.05) when 0.1 ml plasma ultrafiltrate was used for the cytogentic assay (see Materials and Methods), suggests that CF may have an indicator function for HIV disease activity. Decreasing CD4+ T cell numbers predict progression of HIV-1 infection and the CD4+ T cell count is clinically very important for monitoring the disease (42).

Cytokines

A variety of injuries, such as infections, induce a systemic acute-phase reaction. This response is triggered by activation of several stress-sensitive protein kinases, involving reactive oxidants as mediators and leading to synthesis of acute phase cytokines such as TNF-α, IL-1, and IL-2 (43). These cytokines are known to induce cellular production of reactive oxygen species, thereby perpetuating their own formation and action.

A complex network of immunoregulatory cytokines plays an important role in the pathophysiology of HIV-1 infection (44–48). Particularly TNF-α influences the progression of HIV-1 infection by stimulating viral transcription and replication processes, an action synergized by IL-1 and IL-6 (12,49–52). Blood monocytes and macrophages from HIV-infected patients produce high levels of TNF-α (53–55) and IL-6 (55). Gene expression of human immunodeficiency virus depends on host cellular transcription factors, including nuclear transcription factor kappa β (NFκB). NFκB appears to be activated by several different signal transduction pathways (56). Reactive oxygen species have been implicated in NFκB activation, including possibly those generated by TNF-α and IL-1 (57). But recently, a reactive oxidant-independent pathway of NFκB activation has been discovered after stimulation with TNF-α and IL-1 (58,59), and it is believed that activation of NFκB can occur through redox-sensitive as well as redox-insensitive pathways (60).

TNF-α is produced predominantely in response to host injury and infection and has cytotoxic activity against tumor cells. The cellular effects of TNF-α involve generation of oxygen radicals (9,10,61). The net biological effects of TNF-α may often be beneficial to the host because of induction of tolerance to oxidative stress (62). However, overproduction of TNF-α is associated with serious pathological manifestations, including the wasting syndrome associated with neoplastic and infectious diseases, especially that associated with AIDS (63). In our study, TNF-α values were generally within the normal range, but there was a tendency towards elevated TNF-α concentration in HIV-1-negative patients with malignancies (p < 0.1) and in HIV-1-positive patients with malignancies (p < 0.09) in comparison with HIV-1-negative healthy controls.

IL-2 is produced by T and B lymphocytes, promotes proliferation of T cells after antigen stimulation, induces its own synthesis and stimulates growth of natural killer (NK) cells and lymphokine-activated killer (LAK) cells. At high concentrations it can trigger HIV production (64) and induce oxidative stress via stimulated production of reactive oxygen and nitrogen intermediates (11,65). In HIV-1 infection, IL-2 plasma concentration was reported to be decreased (66,67), which is partially a result of CD4+ T cell depletion. Furthermore, the HIV glycoprotein gp 120 suppresses secretion of IL-2 from CD4+ T cells (66). The decrease in IL-2 concentration causes a decline in NK and LAK cell activity, which contributes to the progression of immunosuppression (67). In addition, IL-2 possesses antiviral activity by inducing CD8+ T cells which inhibit HIV-1 production in infected CD4+ T cells (68). These immunostimulatory and anti-HIV activities have led to therapeutical applications of IL-2 in HIV infection (69). Inhibition of HIV reproduction in HIV-infected patients by antiretroviral therapy is associated with a stimulation of IL-2 synthesis and a decreased incidence of opportunistic infections (70,71). In contrast to the reports mentioned above (66,67), in our HIV-1-positive patients, IL-2 plasma levels were in the normal range (Table 1).

IL-6 physiologically promotes B cell growth and differentiation, T cell activation, and cytotoxic T cell differentiation, and it activates NK cells (72,73). IL-6 is synthesized by macrophages, T lymphocytes, fibroblasts, keratinocytes, and other cells. IL-6 concentration was reported to be increased in HIV-1 -positive patients, particularly during the late course of the infection (55,74). The HIV glycoproteins gpl20 and gp 160 induce synthesis of IL-6 in mononuclear cells and CD4+ T cells (75), and HIV-1 tat-protein stimulates IL-6 production in blood monocytes. This may explain increased IL-6 levels in HIV-1-positive patients with advanced disease and high viral replication rates (76). Increased levels of IL-6 during HIV-1 infection exert immunosuppressive effects, such as suppression of T lymphocyte response and inhibition of NK cells (46,76), and it induces HIV-1 expression in infected CD4+ T cells and monocytes (74). High concentrations of IL-6 (55,77) have been reported in HIV-1-positive patients and elevated levels of TNF-α and IL-6 are associated with increased incidence of Kaposi’s sarcoma in HIV-1-infected patients (78). In our study, IL-6 levels were within the physiological range, but they were significantly increased in HIV-1-negative patients with malignancies and in HIV-1-positive patients with asymptomatic disease and zoster, and significantly decreased in HIV patients with malignancies (Kaposi’s sarcoma, lymphomas) in comparison with HIV-1-negative controls (Table 1). The decreased level of IL-6 in our HIV-1-positive patients with malignancies and the normal concentration of IL-2 in our HIV-1-positive patients could be explained by the effects of antiretroviral therapy. The statistically significant negative correlation of IL-6 with CF (r = − 0.43, p < 0.05; Table 2) indicates that IL-6 is not a component of CF in HIV-1-infected patients.

Cytokines such as TNF-α and several inter-leukins exhibit genotoxic properties in human peripheral blood cell cultures (79). The mechanism of these genotoxic effects remains largely unknown. TNF-α, IL-1, and IL-2 are glycopeptides with a molecular weight (m.w.) of 17 kD; the size of IL-6 is 26–32 kD. Thus these molecules pass through the filter YM30, which is used for the ultrafiltration of patients’ plasma. TNF-α is among the identified components of clastogenic factors in vitro. In our study, there was only a tendency towards a positive correlation of CF with TNF-α (r = 0.31, p < 0.14). We have recently shown that 4-hydroxynonenal, which is significantly increased in HIV-1-infected patients (20) and possesses clastogenic activity in vitro (34), tends to correlate (r = 0.39, p < 0.1) with CF of HIV-1-positive patients in vivo (8). The lack of a statistically significant correlation of CF values with TNF-α and 4-hydroxynonenal plasma concentrations in HIV-1 -positive patients may be due to the small numbers of patients studied. However, it may also indicate that the composition of CF may vary from patient to patient, or that other unidentified components contribute to the clastogenic activity of the plasma. Further clinical studies with larger sample populations are necessary to analyze the composition of CF in HIV-1-positive patients.

Acute Phase Proteins

CRP (m.w. 105 kD) is a reliable indicator of acute inflammatory reactions, although it is frequently not elevated in chronic inflammatory conditions. It is synthesized by hepatocytes upon stimulation by IL-6 (80) and IL-1 (81) derived from monocytes/macrophages following tissue disruption and bacterial infection. The physiological function of CRP includes binding of a large variety of glycoproteins and phospholipids present in microorganisms, resulting in inactivation of these organisms. CRP stimulates macrophage functions, such as Chemotaxis, phagocytosis, and oxidative burst, and inhibits neutrophil function. Besides its role as an indicator of inflammatory activity, CRP has cytotropic effects (82) and its anti-tumor effect in animals is possibly mediated via macrophages (83) and involves oxidative pathways. Our finding that CRP is not increased in patients with HIV-1 agrees with those of other reports (13,84–86).

The acute-phase protein neopterin (a pyrazino-pyrimidine derivative) is induced by in-terferon-gamma (IF-γ)-produced from T lymphocytes. Neopterin is released from monocytes/macrophages and its concentration inversely correlates with absolute CD4+ T cell numbers in HIV-1-positive patients, thus it strongly predicts progression of disease from latency to AIDS (14,87). In addition to its role as a sensitive indicator of disease activity in HIV-1 infection, neopterin has been suggested to be associated with the events regulating HIV-1 production and participates in destruction of the cellular immune system (14,87–89). In our study, neopterin levels were significantly increased in HIV-1-negative patients with zoster (p < 0.02) or malignancies (p < 0.03) and in HIV-1-positive patients with zoster (p < 0.05) or malignancies (p < 0.03) (Table 1). Neopterin was not elevated in HIV-1-positive patients with asymptomatic disease. Neopterin (m.w. 253 kD) does not pass through the filter used for the ultrafiltration of patients’ plasma, and there was no significant correlation of neopterin levels with CF activity in patients with HIV, indicating that the two parameters are not related. However, there was a tendency towards a negative correlation of CD4+ T cell numbers with neopterin concentration (r = −0.37, p < 0.1; Table 2), implying an indicator function of neopterin levels in patients with HIV, which confirms the results of Baier-Bitterlich et al. (87).

Plasma levels of granulocyte elastase (30 kD glycoprotein) are increased during a variety of inflammatory reactions and increased release of elastase was found to be associated with HIV infection (90). This increase indicates stimulated neutrophil phagocytosis activity with release of activated oxygen species. TNF-α has been shown to prime neutrophils and macrophages to produce reactive oxidants (91). In our study, TNF-α levels correlated significantly positive with elastase (r = 0.47, p < 0.05), CRP (r = 0.5, p < 0.02), and neopterin (r = 0.44, p < 0.05) (Table 2). Several studies have shown that increased elastase concentrations in patients’ plasma indicate serious complications (92–94). In our study there was a tendency towards increased elastase levels only in HIV-positive patients with zoster (p < 0.1), which indicates that most of our patients were vitally not endangered. There was no significant correlation of elastase concentration with CF activity in the patients with HIV (data not shown), indicating that the two parameters are not related.

Although CRP and elastase were not significantly increased in our patients, CRP correlated positively with elastase (r = 0.92, p < 0.002) and neopterin (r = 0.79, p < 0.002), and elastase correlated positively with neopterin (r = 0.72, p < 0.002). This highly significant correlation indicates that the acute-phase proteins are useful as inflammatory markers, but their response differs in specificity and sensitivity.

Conclusions

CFs were significantly increased in all HIV-1-positive patients. The highest CF values were observed in the HIV patients with Kaposi’s sarcoma. CFs are not induced by viral infections in general, as they were not elevated in HIV-negative zoster patients. CFs are not specific for HIV infection, as they are significantly elevated in HIV-negative patients with malignancies. Although several cytokines have genotoxic properties in vitro, there was a tendency towards a positive correlation between TNF-α plasma concentration and CF values only in HIV patients. The lack of correlation of CF with IL-6 and neopterin indicates that these factors do not contribute to the clastogenic activity of the HIV patient’s plasma. The composition of CF may vary from patient to patient and/or other unidentified substances may contribute to the CF activity of HIV-1-infected patients. Since the formation and the action of clastogenic factors are modulated by superoxide anion radicals, we suggest that the superoxide-scavenging activity of the plasma is an important parameter that determines the expression of clastogenic activity. CFs are sensitive and long-lived markers of disturbed pro-oxidant/antioxidant balance. They can be determined readily by cytogenetic analysis and may be useful for evaluation of drugs and other intervention strategies in HIV infection.

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Acknowledgements

The assistance of Dr. Ackermann, Institute of Biomathematics, J. W. Goethe University Medical School, Frankfurt, in calculating the statistical analysis is highly appreciated.

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Authors and Affiliations

  1. Department of Dermatology, Frankfurt University Hospital, Frankfurt am Main, Germany

    Jürgen Fuchs (Privatdozent), Nicole Oelke, Matthias Imhof, Falk Ochsendorf & Helmut Schöfer

  2. Department of Internal Medicine, Frankfurt University Hospital, Frankfurt am Main, Germany

    Gerhard Oromek

  3. Institut Biomedical des Cordeliers, Université Pierre et Marie Curie, Paris, France

    Amina Alaoui-Youssefi & Ingrid Emerit

  4. Heinsestrasse 8, 63739, Aschaffenburg, Germany

    Jürgen Fuchs (Privatdozent)

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Fuchs, J., Oelke, N., Imhof, M. et al. Multiparameter Analysis of Clastogenic Factors, Pro-oxidant Cytokines, and Inflammatory Markers in HIV-1-Infected Patients with Asymptomatic Disease, Opportunistic Infections, and Malignancies. Mol Med 4, 333–343 (1998). https://doi.org/10.1007/BF03401740

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