Potential pathogenetic role of Th17, Th0, and Th2 cells in erosive and reticular oral lichen planus.

OBJECTIVES: The role of Th17 cells and associated cytokines was investigated in oral lichen planus. MATERIAL AND METHODS: 14 consecutive patients with oral lichen planus were investigated. For biological studies, tissues were taken from reticular or erosive lesions and from normal oral mucosa (controls) of the same patient. mRNA expression for IL-17F, IL-17A, MCP-1, IL-13, IL-2, IL-10, IL-1ß, RANTES, IL-4, IL-12B, IL-8, IFN-γ, TNF-α, IL-1α, IL-18, TGF-ß1, IL-23R, IL-7, IL-15, IL-6, MIG, IP-10, LTB, VEGF, IL-5, IL-27, IL-23A, GAPDH, PPIB, Foxp3, GATA3, and RORC was measured using the QuantiGene 2.0. RESULTS: Results showed that Th17-type and Th0-type molecules' mRNAs, when compared with results obtained from tissue controls, were increased in biopsies of erosive lesions, whereas Th2-type molecules' mRNAs were increased in reticular lesions. When the CD4+ T-cell clones, derived from oral lichen planus tissues and tissue controls, were analyzed, a higher prevalence of Th17 (confirmed by an increased CD161 expression) and Th0 CD4+ T clones was found in erosive lesions, whereas a prevalence of Th2 clones was observed in reticular lesions. CONCLUSIONS: Our data suggest that Th17, Th0, and Th2 cells, respectively, may have a role in the pathogenesis of erosive and reticular oral lichen planus.

Natural killer cell deficiencies in a consecutive series of children with herpetic encephalitis.

Natural killer (NK) cells play a fundamental role in innate and early phases of adaptive immunity against viral infections, both in humans and in animal models. To date, NK cell deficiencies in patients with severe herpetic infections have been reported in single cases, and their role as predisposing factor is still controversial. Five children affected by herpetic encephalitis were consecutively admitted to the Anna Meyer Children's Hospital in Florence (Italy) between 2003 and 2005. We therefore investigated the presence of NK cell deficiencies in a consecutive series of children with herpetic encephalitis. Five healthy children were included in the study as controls. Differential WBC counts, main Ig and IgE class serum analysis, cytofluorimetric analysis of circulating T, B and NK cells were performed on our study population. Sequencing of a selected region of CD16A gene transcript was carried out in two patients. All patients resulted to be affected by deficiencies related to NK cells in respect to controls. One patient was also affected by lymphopenia, while no other significant deficits of immunity were detected in the study population. To date, this is the first survey that demonstrates isolated NK cell deficiencies in a cohort of consecutive patients affected by severe herpes simplex infections. These findings suggest a role for NK cell deficiencies as a predisposing factor for increased susceptibility and severe course of disease in these patients.

Influence of impaired T- and B-cell compartments on efficacy of IVIg in dysimmune neuropathies.

Autoimmune mechanisms are postulated to play a role in the development and progression of dysimmune neuropathies (DN). We investigated the relation between lymphocyte number and marker expression, and disease activity in 20 patients with DN under intravenous immunoglobulins (IVIg) treatment. B- and T-lymphocyte markers were studied by flow cytometry of the expression of CD5, CD25, CD23 and CD38 markers on B cells and of CD3, CD4 and CD8 markers, respectively. These parameters were compared with those obtained from matched healthy volunteers. The proportions of CD38+ B cells were higher in patients compared with those of controls. Proportions of activated CD4+ and CD8+ T cells were comparable in peripheral blood mononuclear cells of patients and controls, but a significant reduction of the absolute numbers of CD3+, CD4+ and CD8+ cells were observed in DN patients. The percentages of CD25+ memory T cells were instead significantly increased in DN patients. Lastly, T-cell reduction and the CD19/CD38 ratio over total B (CD19+) cells directly correlated with a poor response to IVIg therapy. In DN, whereas T-cell number is reduced, activated T and B cells are increased, thus suggesting an intrinsic defect of the immune response.

Embryonic soluble HLA-G as a marker of developmental potential in embryos.

BACKGROUND: In human reproduction, embryo implantation is complex and poorly understood. At present, no single markers are used in routine treatment to assay biochemical functions of the human embryo. Soluble human leukocyte antigen-G (sHLA-G) could be considered a possible marker of embryo developmental potential. It is localized primarily on the extravillous trophoblast, making this antigen a potential mediator of immune interaction at the maternal-fetal interface during gestation. METHODS: Soluble-HLA-G levels were evaluated by an enzyme-linked immunosorbent assay (ELISA) employing monoclonal antibody MEM-G9. It was evaluated in 318 media of single embryo cultures. We correlated the presence of sHLA-G with embryo morphology and the pregnancy obtained in that treatment cycle. RESULTS: No correlation was found between embryo morphology and sHLA-G levels. Pregnancy was observed only when the medium of at least one transferred embryo contained sHLA-G. In 26 out of 66 patients, none of the obtained embryos showed any detectable sHLA-G molecules and no pregnancy occurred. CONCLUSIONS: From our results, we propose sHLA-G as a potential marker of embryo development: the sHLA-G ELISA can be a useful biochemical assay in addition to embryo morphology in embryo selection for transfer in IVF treatment if there are other embryos with the same morphology.

Lectin histochemistry in the human placenta of pregnancies complicated by intrauterine growth retardation based on absent or reversed diastolic flow.

The oligosaccharide distribution of the glycoconjugates was investigated in placental tissue of pregnancies complicated by intrauterine growth retardation (IUGR) with absent or reversed flow in the umbilical artery (ARED), between the 29 and the 37 weeks of pregnancy. Placentae of a gestational age-matched group of normally grown pregnancies was also selected as control group. For this purpose a battery of seven HRP-conjugated lectins was used (DBA, SBA, PNA, ConA, WGA, LTA and UEA I). Our data showed that alpha-D-mannose (ConA), N-acetyl-D-glucosamine (WGA), beta-N-acetyl-D-galactosamine (SBA), alpha-L-fucose (LTA and UEA I) were present in less amount or were not present in the trophoblast and/or in the endothelial cells of the pathological group compared to the control one. The trophoblast basement membrane and/or basal plasma membrane of the pathological placentae were characterized by the presence of alpha-D-mannose (ConA), N-acetyl-D-glucosamine (WGA), sialic acid and D-galactose-(beta1-->3)-N-acetyl-D-galactosamine (neuraminidase-PNA), only in some tracts, in all the weeks of gestation. In the control placentae these sugar residues were present in the whole basement membrane and/or basal plasma membrane from 31 or 35 weeks. The Hofbauer cells of the pathological placental tissue showed a less amount or lack of alpha-D-mannose (ConA), beta-N-acetyl-D-galactosamine (SBA) and alpha-L-fucose (UEA I) compared to the control ones. These results suggest that a less amount or lack of some sugar residues may contribute to restricted placenta growth and development and thus reduced efficiency in maternal-fetal exchanges of gases and metabolites.

Influence of maternal magnesium sulphate and ritodrine treatment on cerebral blood flow velocity of the preterm newborn.

BACKGROUND: To evaluate the effect of antenatal tocolytic administration of magnesium sulphate and ritodrine on the cerebral blood flow velocity and on the cerebral vascular resistance of preterm newborns in the first hours of life. METHODS: Cerebral blood flow velocity, resistance index and relative vascular resistance were studied in 27 preterm infants (<34 weeks gestation) with antenatal exposure to maternal magnesium sulphate treatment and in 27 preterm infants (<34 weeks gestation) with antenatal exposure to maternal ritodrine treatment. Both antenatal magnesium sulphate or ritodrine were used for tocolysis. Cerebral blood flow was measured, using Doppler ultrasonography, in the anterior cerebral artery, in the left middle cerebral artery and in the right middle cerebral artery. RESULTS: We did not find any significant difference in the blood flow velocity, resistance index or relative vascular resistance in the three cerebral arteries between the two treatment groups. CONCLUSIONS: Our study shows that maternal antenatal administration of magnesium sulphate to delay preterm delivery, compared to antenatal administration of ritodrine, does not induce any significant differences either in cerebral blood flow velocity or in cerebral vascular resistance of preterm infants in the first hours of life.

Functional and ultrastructural features of DNA-fragmented human sperm.

The functional significance of deoxyribonucleic acid (DNA) fragmentation in ejaculated human sperm is unclear. In this study the extent of DNA strand breakage in swim-up selected spermatozoa was evaluated by terminal deoxynucleotidyl transferase-mediated fluorescein-dUTP nick end labeling (TUNEL)-coupled flow cytometry and correlated with several functional and morphological sperm parameters. The extent of DNA fragmentation (mean = 11.07%+/-8.00%, range = 0.79%-42.64%, n = 140) was positively related to abnormal morphology and associated with defects of the sperm tail. A negative correlation was found between DNA breakage and progressive motility. When a stepwise multiple linear regression model was used to analyze the relationship between DNA fragmentation and the aforementioned parameters, only motility results were included in the model. The presence of spermatozoa showing submicroscopic characteristics resembling those of somatic apoptosis has been reported in human ejaculate. To verify whether sperm DNA fragmentation was associated with the presence of such apoptotic-like cells, we performed electron microscopy and TUNEL-coupled flow cytometry in a limited number of sperm samples (n = 24). Although we did not observe any significant relationship between DNA breakage and the characteristics that are suggestive of apoptosis, an association was found with several ultrastructural features, indicating an impaired motility. Hence, we conclude that in ejaculated sperm, DNA fragmentation does not correspond to the apoptosis-like phenomenon and that it is associated with defects of motility.

Effects of low day 3 luteinizing hormone levels on in vitro fertilization treatment outcome.

In a previous study we demonstrated that women with day 3 luteinizing hormone (LH) values < 3 IU/l subjected to controlled ovarian hyperstimulation without pituitary desensitization responded with a lower number of follicles > 15 mm compared to women with a higher basal LH level. The aim of this study was to determine whether in patients with day 3 LH levels < 3 IU/l a further reduction of serum LH concentration by gonadotropin-releasing hormone (GnRH) analog impairs follicular response to follicle stimulating hormone (FSH) and treatment outcome in in vitro fertilization (IVF) cycles. For this purpose we retrospectively studied 249 consecutive women subjected to standard IVF treatment employing pituitary desensitization with buserelin and follicular stimulation with urinary highly purified FSH. The patients were divided into two groups according to their day 3 LH value. The first group (group A) showed day 3 LH levels < 3 IU/l and the second (group B) had day 3 LH levels > 3 IU/l. Group A and B patients did not show statistically significant differences in the ovarian response to FSH, nor in IVF treatment outcome, showing that in FSH treated GnRH analog suppressed cycles, the ovarian responsiveness and IVF outcome do not differ according to basal LH values. However, the high dosage of FSH we employed in group A and B patients could account, at least in part, for this result. Indeed, comparative evaluations with unsuppressed cycles (our previous study) strongly suggest that a reduced ovarian responsiveness to gonadotropins in patients with day 3 LH values < 3 IU/l should be considered in clinical practice.

Changes in mesenteric blood flow response to feeding: conventional versus fiber-optic phototherapy.

OBJECTIVE: To evaluate whether fiberoptic phototherapy influences the postprandial increase in mesenteric blood flow velocity similarly to conventional phototherapy in preterm infants. PATIENTS AND METHODS: With the use of Doppler color ultrasonography, blood flow velocity in the superior mesenteric artery was measured both preprandially and postprandially in 19 preterm infants during and after conventional phototherapy, and in 20 preterm infants during and after fiber-optic phototherapy. The mean arterial blood pressure/mean flow velocity ratio was calculated as an estimate of relative vascular resistance of the superior mesenteric artery. RESULTS: The study shows that conventional phototherapy blunts the postprandial mesenteric blood flow response to feeding in preterm infants. Furthermore, it shows that the postprandial increase in intestinal blood flow is not attenuated when fiber-optic phototherapy is administered, and that such postprandial increase of blood flow is significantly greater than in infants receiving conventional phototherapy. During and after fiber-optic phototherapy, a significant reduction in postprandial relative vascular resistance was found; such reduction was significantly greater than during conventional phototherapy. CONCLUSIONS: Fiber-optic phototherapy is preferable to conventional phototherapy for the treatment of hyperbilirubinemia in preterm infants because it does not affect the physiologic postprandial redistribution of blood flow from the periphery to the gastrointestinal system as does conventional phototherapy.

Effects of indomethacin and ibuprofen on mesenteric and renal blood flow in preterm infants with patent ductus arteriosus.

OBJECTIVE: To evaluate the effect of intravenous ibuprofen and indomethacin for treatment of patent ductus arteriosus (PDA) on mesenteric and renal blood flow velocity in preterm infants. STUDY DESIGN: Seventeen mechanically ventilated preterm infants (<33 weeks' gestation) with PDA received either 0.2 mg/kg indomethacin (n = 8) or 10 mg/kg ibuprofen (n = 9), infused over 15 minutes. Mesenteric and renal blood flow velocity were measured by using Doppler ultrasonography. RESULTS: Indomethacin caused a significant reduction in mesenteric and renal blood flow velocity 30 minutes after drug administration; mesenteric and renal blood flow velocity did not return to the pretreatment values by 120 minutes. Ibuprofen did not alter blood flow 30 minutes after treatment, and blood flow increased 120 minutes after treatment. Mesenteric and renal blood flow velocity changes were significantly different between the 2 treatment groups. CONCLUSIONS: Compared with indomethacin, ibuprofen did not significantly reduce mesenteric and renal blood flow velocity.

Serum CA-125 values on the day of oocyte retrieval are not predictive of subsequent pregnancy with in-vitro fertilization.

In the clinical management of in-vitro fertilization (IVF) patients it would be very useful to know, before the embryo transfer, whether or not there is a significant chance of pregnancy in that cycle. If low, it would be better to freeze the embryos and postpone the embryo transfer to a subsequent cycle. For this reason, a retrospective study was carried out to investigate the correlations between the serum CA-125 values before embryo transfer and the clinical outcome of that IVF cycle. Women aged <40 years undergoing a complete infertility evaluation including laparoscopy and receiving gonadotrophin-releasing hormone analogue (GnRHa) suppression followed by purified follicle stimulating hormone (FSH) for IVF-embryo transfer were entered into the study. Ninety-seven cycles qualified for evaluation (26 pregnant and 71 non-pregnant cycles). CA-125 concentrations on the day of oocyte retrieval were significantly lower in the pregnant versus non-pregnant cycles in both non-endometriosis and endometriosis patients. To evaluate the existence of a cut-off value of CA-125 which would allow the prediction of a possible pregnancy with sufficient specificity and sensitivity, a receiver operating characteristic curve analysis was performed. This analysis demonstrated the absence of any predictive value of the subsequent pregnancy for CA-125 concentrations. For this reason, and in contrast with previous findings, CA-125 determinations before the embryo transfer in IVF patients do not appear to be a useful tool for clinicians to use in predicting the outcome of IVF in any given cycle.

Predicting ovarian malignancy: application of artificial neural networks to transvaginal and color Doppler flow US.

PURPOSE: To compare the performance of artificial neural networks (ANNs) with that of multiple logistic regression (MLR) models for predicting ovarian malignancy in patients with adnexal masses by using transvaginal B-mode and color Doppler flow ultrasonography (US). MATERIALS AND METHODS: A total of 226 adnexal masses were examined before surgery: Fifty-one were malignant and 175 were benign. The data were divided into training and testing subsets by using a "leave n out method." The training subsets were used to compute the optimum MLR equations and to train the ANNs. The cross-validation subsets were used to estimate the performance of each of the two models in predicting ovarian malignancy. RESULTS: At testing, three-layer back-propagation networks, based on the same input variables selected by using MLR (i.e., women's ages, papillary projections, random echogenicity, peak systolic velocity, and resistance index), had a significantly higher sensitivity than did MLR (96% vs 84%; McNemar test, p = .04). The Brier scores for ANNs were significantly lower than those calculated for MLR (Student t test for paired samples, P = .004). CONCLUSION: ANNs might have potential for categorizing adnexal masses as either malignant or benign on the basis of multiple variables related to demographic and US features.

Frequency of a mutated CCR-5 allele (delta32) among Italian healthy donors and individuals at risk of parenteral HIV infection.

The aim of this study was to assess the frequency of a truncated allele of the CCR-5 gene (delta32) in Italy, and address its possible role in parenteral HIV transmission, as well as its influence in HIV-associated disease progression. In 371 unrelated seronegative healthy blood donors the delta32 allele frequency was 0.047; this figure was significantly different from those reported in northern America and northern Europe populations. However, delta32 allele frequency in healthy individuals did not differ significantly from that found in 54 seronegative drug users (0.065), 98 seronegative hemophiliacs (0.051), and 81 seropositive hemophiliacs (0.049). Although in seropositive hemophiliacs the wt/delta32 heterozygous genotype was associated with a trend to a slower decline in CD4+ cell counts, its presence did not seem to influence disease progression, as comparable delta32 allele frequency frequencies were found among progressing (0.042) and nonprogressing (0.111) patients. These data do not seem to support a protective role of the delta32 allele in preventing HIV infection through the parenteral route, or in influencing the natural history of the disease in this particular risk category, although the delta32 heterozygous state was associated with lower plasma viremia levels. On the other hand, the finding of non-syncytium-inducing HIV strains in the majority of delta32 heterozygous seropositive patients suggests that its presence could not be a major factor in driving a switch toward more cytopathic, T-tropic HIV strains through selective pressure in coreceptor usage.

Placental morphometry in pregnancies complicated by intrauterine growth retardation with absent or reversed end diastolic flow in the umbilical artery.

The aim of this study was to assess any possible correlation between villous tree architecture and its vascularization, and absent or reversed end-diastolic flow velocity (ARED) in the umbilical artery. The study group included seven pregnancies complicated by IUGR (estimated fetal weight < 10th percentile) and absent end-diastolic flow velocity in the umbilical artery. A gestational-age matched group of seven normally grown pregnancies was selected as control group. At delivery, the placenta was weighed and immersed in 10% neutral buffered formalin. A stratified random sampling procedure was used to obtain 12 blocks of full-thickness tissue per organ. A single random section was cut from each block. The following morphometric parameters were evaluated in each section: mean vessel diameter, volume density of the villous tissue, stem villi and terminal villi. Measurements were performed using a computerized Video Image Analysis system. No significant difference in mean vessel diameter was found between the two groups (37.1 microns versus 36.1 microns; p = 0.1). There was a significant reduction in the proportion of total villous tissue in the ARED group (43% versus 52%): this was due to a significant reduction in the volume of tissue occupied by the terminal villi (14.1% versus 18.4%). No significant difference was found in the proportion of villous tissue occupied by the stem villi (42% versus 40%). Several studies have investigated the anatomical and/or vasomotor bases of absent end diastolic flow velocity in the umbilical artery of fetuses with severe IUGR. Our observations of a significant reduction in the proportion of villous tissue occupied by the peripheral villi are consistent with the theory that failure of normal development of the terminal villous is responsible for the increased vascular resistance in IUGR pregnancies with ARED.

Maternal serum screening for trisomy 18 in the first trimester of pregnancy.

The aim of this study was to evaluate the feasibility of first-trimester biochemical screening for trisomy 18, by using pregnancy-associated plasma protein A (PAPP-A) and free beta human chorionic gonadotropin (hCG) in combination with maternal age. Maternal serum levels of PAPP-A and free beta-hCG were assayed retrospectively in stored sera from 323 singleton pregnancies at 8-13 weeks' gestation. These samples included 23 trisomy 18 pregnancies and 300 unaffected controls, diagnosed either at chorionic villus sampling or at mid-trimester amniocentesis. The median MOM in affected pregnancies was 0.25 for PAPP-A and 0.34 for free beta-hCG. Statistically significant reductions were found in the mean levels of both PAPP-A (t test: P < 0.000001) and free beta-hCG (p < 0.000001) in trisomy 18 affected pregnancies when compared with the unaffected samples. Screening for trisomy 18 using a combination of maternal age, PAPP-A and free beta-hCG would achieve a detection rate of 76.6 per cent for a false-positive rate of 0.5 per cent. These results suggest that first-trimester biochemical screening for trisomy 18 might be possible. Further investigations in a general population are needed before introducing such screening into clinical practice.

First trimester screening for Down's syndrome using maternal serum PAPP-A and free beta-hCG in combination with fetal nuchal translucency thickness.

The aim of this study was to evaluate the potential effectiveness of maternal serum pregnancy-associated plasma protein A (PAPP-A) and free beta-hCG in combination with nuchal translucency thickness in first trimester screening for Down's syndrome. Maternal serum levels of PAPP-A and free beta-hCG were assayed in stored sera from 32 Down's syndrome and 200 unaffected pregnancies. Fetal nuchal translucency was measured by ultrasound at the time of blood sampling. Screening of Down's syndrome using a combination of maternal age, PAPP-A, free beta-hCG and nuchal translucency would achieve a detection rate of 75.8% for a false positive rate of 5%.

Anatomical-radiological and morphometric analysis of the lateral atlantoaxial joint.

The width of the articular space of the lateral atlantoaxial joint was measured to evaluate normal range values and possible variations according to age and sex. The study was based on X-rays of the lateral atlantoaxial joint of 52 females (age range: 18-82 ys) and 50 males (age range: 20-82 ys). The radiograms were taken using a semiautomatic computerized morphometric system (Videoplan II, Image Analysis system-Kontron, Munich, Germany). The articular space width of the lateral atlantoaxial joint was measured on both sides, at the medial, lateral and median levels. The mean value at these levels, on both sides, was calculated. Regression analysis was performed to evaluate the relationships between the width of the articular space at the three different levels and the patient's age and gender. No significant differences were found in the mean values of the articular space width at the three levels as regards gender. However, a significant inverse correlation was found between the articular space width and the patients' age in both males and females. Comparative analysis of regression lines showed that the slope of these lines was not significantly different when the sexes were compared. On the contrary, the intercept of the regression lines at the medial and median levels was significantly higher in the females than in the males; the contrary was encountered at the lateral level. This feature is probably due to the different types of inclination of the articular facets of the axis or of those of the atlas in the two sexes.

Low day 3 luteinizing hormone values are predictive of reduced response to ovarian stimulation.

The aim of the present work was to evaluate whether low day 3 luteinizing hormone (LH) values in the presence of normal follicle stimulating hormone (FSH) are predictive of poor response to ovarian stimulation. Two groups of women undergoing ovarian stimulation and differing only in the day 3 LH concentration (<3 mIU/ml, study group, n=30; >3 mIU/ml, control group, n=45) were retrospectively analysed. Study group patients developed a lower oestradiol peak (703+/-388 versus 955+/-400 ng/ml; P = 0.005) and a lower number of follicles >15 mm diameter at the time of human chorionic gonadotrophin (HCG) administration (2.6+/-1.3 versus 3.6+/-1.8; P=0.004) than the control group. Conversely, a similar ratio of oestradiol: follicles >15 mm diameter was observed (256+/-118 versus 269+/-93; P=0.563). The number of follicles >10 mm at the time of HCG administration appeared to be lower in the study group, but this difference was not statistically significant (6+/-3.9 versus 7.8+/-4.3). Our data indicate that day 3 LH values <3 mIU/ml are predictive of poor response to ovarian stimulation.

Functional characterization and modulation of cytokine production by CD8+ T cells from human immunodeficiency virus-infected individuals.

CD8+ T-cell clones were generated from peripheral blood mononuclear cells (PBMC) of three human immunodeficiency virus (HIV)-seronegative individuals and six HIV-seropositive individuals and assessed for their cytokine secretion profile, cytolytic potential, and chemokine production. While the great majority of CD8+ T-cell clones generated from HIV-seronegative individuals produced interferon (IFN)-gamma, but not interleukin-4 (IL-4), that is a type 1 cytotoxic (Tc1) profile, high numbers of CD8+ T-cell clones generated from HIV-seropositive individuals produced IL-4 in addition to IFN-gamma or IL-4 alone, thus showing a type 0 cytotoxic (Tc0)- or a type 2 cytotoxic (Tc2) profile, respectively. Tc0/Tc2 cells displayed lower cytolytic activity than Tc1 cells, including a reduced ability to lyse autologous targets pulsed with HIV or HIV peptides. By contrast, the production of chemokines RANTES and macrophage inflammatory protein-1alpha was comparable in Tc1, Tc0, and Tc2 clones irrespective of whether they were derived from HIV-seronegative or HIV-seropositive individuals. When CD8+ T-cell clones were generated from PBMC cultures of HIV-seronegative individuals conditioned with IL-4 plus an anti-IL-12 antibody (Ab), a shift towards the Tc0/Tc2-like profile was observed. Conversely, the addition to PBMC cultures of IL-12 plus an anti-IL-4 Ab shifted the differentiation of CD8+ T cells from HIV-infected individuals towards the Tc1-like profile, whereas IL-12 or anti-IL-4 Ab alone had a lower Tc1-promoting effect. Irradiated PBMC from HIV-infected individuals, used as feeder cells, shifted the differentiation of CD8+ T cells from a healthy HIV-seronegative individual towards the Tc0/Tc2-like profile. On the other hand, a shift towards the Tcl-like profile was noted in CD8+ T-cell clones generated from the skin specimens of two HIV-seropositive patients with Kaposi's sarcoma, successfully treated with IFN-alpha, in comparison to CD8+ clones generated from the same skin areas before treatment. The IFN-alpha-induced Tc1 shift could be prevented by the incubation of skin-infiltrating CD8+ T cells with IL-4 before cloning. Taken together, these data indicate that both defective production of IL-12 and abnormal IL-4 production in bulk PBMC populations of HIV-infected individuals may contribute to the development of high numbers of CD8+ T-cell clones showing a Tc0/Tc2-like phenotype and reduced cytolytic potential against HIV itself. They also suggest that the cytokine profile of CD8+ T-cell clones can be modulated by cytokines (or anticytokine Ab) both in vitro and in vivo.