Efficacy of topical imiquimod 3.75% in the treatment of actinic keratosis of the scalp in immunosuppressed patients: our case series.

Background: Actinic keratoses (AK) represent common cutaneous lesions, appearing in 'Field cancerization areas' and potentially evolving toward invasive neoplasm. Immunosuppressed patients frequently develop numerous and aggressive AKs.Aim: In this observational study, we report our experience with topical Imiquimod 3.75% as 'Field-directed therapy' in a cohort of immunosuppressed patients.Methods: A group of 13 immunosuppressed patients presenting multiple AKs of the balding scalp was treated with topical Imiquimod 3.75%. Each patient underwent clinical examination at fixed timepoints during the treatment (T0, T14, T28, T42) and eight weeks after the end.Results: In our cohort, the treatment was well tolerated, with minimal local adverse events. We observed a good clinical response, in terms of Lmax lesions (maximum lesion count during the course of therapy) and of AK clearance. In our group, 46% of patients showed no detectable lesions at the end of the observation period, and this result was maintained up to 1 year after the end of treatment.Conclusion: Topical Imiquimod 3.75% represents an effective and safe treatment for multiple AK of the scalp also in immunosuppressed patients. To the best of our knowledge, this is the first report on the use of this drug in this category of subjects.

Antineoplastic activity of idazoxan hydrochloride.

PURPOSE: Idazoxan hydrochloride (IDA) is a 241 molecular weight imidazoline and adrenoreceptor ligand. It binds to mitochondrial membranes and promotes apoptosis of pancreatic beta cells. Since IDA has not been tested against tumor cells, the purpose of our study was to determine if IDA has antineoplastic activity. METHODS: We used the conversion of a soluble tetrazolium salt to an insoluble formazan precipitate and differential staining cytotoxicity assays to determine if IDA was cytotoxic to cell lines of murine lung cancer and human prostate cancer, as well as to a variety of fresh human tumor samples. We used flow cytometry to analyze cell death and calreticulin expression. RESULTS: IDA is cytotoxic to both cell lines and against aliquots of specimens of breast, gastric, lung, ovarian and prostate cancers as well as non-Hodgkin's lymphoma. It produces apoptotic cell death and promotes calreticulin expression, suggesting that IDA might be immunomodulatory in vivo. CONCLUSION: We anticipate that IDA will be clinically useful in cancer treatment.

Antibody from patients with acute human immunodeficiency virus (HIV) infection inhibits primary strains of HIV type 1 in the presence of natural-killer effector cells.

The partial control of viremia during acute human immunodeficiency virus type 1 (HIV-1) infection is accompanied by an HIV-1-specific cytotoxic T-lymphocyte (CTL) response and an absent or infrequent neutralizing antibody response. The control of HIV-1 viremia has thus been attributed primarily, if not exclusively, to CTL activity. In this study, the role of antibody in controlling viremia was investigated by measuring the ability of plasma or immunoglobulin G from acutely infected patients to inhibit primary strains of HIV-1 in the presence of natural-killer (NK) effector cells. Antibody that inhibits virus when combined with effector cells was present in the majority of patients within days or weeks after onset of symptoms of acute infection. Furthermore, the magnitude of this effector cell-mediated antiviral antibody response was inversely associated with plasma viremia level, and both autologous and heterologous HIV-1 strains were inhibited. Finally, antibody from acutely infected patients likely reduced HIV-1 yield in vitro both by mediating effector cell lysis of target cells expressing HIV-1 glycoproteins and by augmenting the release of beta-chemokines from NK cells. HIV-1-specific antibody may be an important contributor to the early control of HIV viremia.

Osteoclasts formed by measles virus-infected osteoclast precursors from hCD46 transgenic mice express characteristics of pagetic osteoclasts.

Pagetic osteoclasts (OCLs) are abnormal in size and contain paramyxoviral-like nuclear inclusions that cross-react with antibodies to measles virus (MV). However, the role that MV infection plays in Paget's disease is unknown, because no animal model of Paget's disease is available. Therefore, we targeted a cellular MV receptor, human CD46 (hCD46), to cells in the OCL lineage in transgenic mice using the mouse tartrate-resistant acid phosphatase (TRAP) gene promoter. In vitro infection of OCL precursors from hCD46 transgenic mice with MV significantly increased OCL formation in bone marrow cultures. The numbers of TRAP-positive mononuclear cells and CFU-GM, the earliest identifiable OCL precursor, were also significantly increased. MV-infected OCLs formed from hCD46 marrow were increased in size, contained markedly increased numbers of nuclei, and had increased bone-resorbing capacity per OCL compared with OCLs formed from marrow of nontransgenic littermates. Furthermore, IL-6 and 24-hydroxylase messenger RNA expression levels were increased in MV-infected hCD46 transgenic mouse bone marrow cultures. Treatment of MV-infected hCD46 marrow cultures with a neutralizing antibody to IL-6 blocked the increased OCL formation seen in these cultures. These data demonstrate that MV infection of OCL precursors results in OCLs that have many features of pagetic OCLs, that the enhanced OCL formation is in part mediated by increased IL-6 expression induced by MV infection, and suggest that the hCD46 transgenic mouse may be a useful model for examining the effects of MV infection on OCL formation in vivo.

Relationship between antibody-dependent cellular cytotoxicity, plasma HIV type 1 RNA, and CD4+ lymphocyte count.

To explore the role of antibody-dependent cellular cytotoxicity (ADCC) in controlling HIV-1 infection, ADCC was compared with plasma RNA and CD4+ cell count in 40 patients not receiving antiretroviral therapy and in seven patients after the initiation of treatment. Among untreated patients, ADCC effector cell function of peripheral blood mononuclear cells, measured by (51)Cr release assay, correlated inversely with viral load (R = -0.42, p = 0.007) and directly with CD4+ cell count (R = 0.52, p = 0.001). On the other hand, HIV-1-specific ADCC antibody level correlated directly with viral load, but only among patients with high CD4+ cell counts. Therapy-induced changes in ADCC effector cell function correlated strongly with changes in CD4+ cell count (R = 0.86, p = 0.014), whereas there was no consistent pattern of change in ADCC antibody with therapy. In a novel assay, ADCC reduced virus yield from CD4+ lymphocytes infected with a primary HIV isolate. ADCC may contribute to control of viremia, and CD4+ lymphocytes likely play a role in ADCC effector and antibody functions.

Antibody inhibition of cytomegalovirus: the role of natural killer and macrophage effector cells.

To explore mechanisms by which antibody might inhibit cytomegalovirus (CMV), we measured the ability of intravenous CMV-IgG (CytoGam) to reduce viral yield in the presence of effector cells. Foreskin fibroblasts were infected with a clinical strain of CMV, and CytoGam was added along with effector cells consisting of either unfractionated peripheral blood mononuclear cells (PBMCs), natural killer (NK) cells, or macrophages. The combination of CytoGam and either of the effector cell types markedly inhibited established CMV infection in vitro. In addition, CytoGam combined with effector cells protected the monolayer from CMV-induced cytopathic effects. Antibody-dependent, effector cell-mediated functions may underlie the ability of CytoGam to prevent or modulate CMV infection in vivo.

Sex-associated differences in the antibody-dependent cellular cytotoxicity antibody response to measles vaccines.

In some countries, excessive non-measles-related mortality has been observed among female recipients of high-titer measles vaccines. We determined if differences in the immune response to measles vaccines underlie the excessive female mortality by measuring the measles virus (MV)-specific antibody-dependent cellular cytotoxicity (ADCC) antibody response in 65 3-year-old Gambian children immunized with Edmonston-Zagreb medium-titer (EZ) or Schwarz standard vaccines during infancy. Among the 20 females and 22 males with undetectable anti-MV antibodies at the time of immunization, females had significantly lower ADCC than males (median cytotoxicities of 1/100 serum dilutions = 8.4 and 12%, respectively; P = 0.04). This sex-associated difference was present only among the six female and seven male recipients of EZ vaccine (median cytotoxicities = 5.1 and 19.0%, respectively; P = 0.02). There were no significant sex-associated differences in neutralizing antibody activity. Decreased ADCC antibody activity may contribute to the lower survival rate observed in females receiving high-titer measles vaccination.

Antibody-dependent cellular cytotoxicity independently predicts survival in severely immunocompromised human immunodeficiency virus-infected patients.

The exact immune defects leading to human immunodeficiency virus (HIV)-associated opportunistic infections, malignancies, and death are unknown. In this study, the relationship between survival and 2 immune functions, cytomegalovirus-specific antibody-dependent cellular cytotoxicity (ADCC) and natural killer (NK) activity, was determined by using peripheral blood mononuclear cells from 39 severely immunocompromised patients (median CD4 count, 7). Median follow-up was 414 days; 15 subjects died and 24 remained alive. In a Kaplan-Meier analysis, high baseline ADCC (>median) was associated with improved survival (P=.05). A similar trend was observed for NK activity (P=.1). In a multivariate model controlling for baseline CD4 cell count, HIV RNA, and use of protease inhibitors during follow-up, high ADCC, but not high NK activity, remained significantly associated with a lower risk of death (relative risk, 0.18; 95% confidence interval, 0.05-0.75). ADCC may be an important determinant of disease progression independently of anti-retroviral therapy, CD4 cell count, and HIV RNA.

In vitro reduction of virus infectivity by antibody-dependent cell-mediated immunity.

Antibody-dependent cellular cytotoxicity (ADCC), an important defense against viral infections, is generally measured in 51Cr-release assays. However, the effect of ADCC on viral burden is more relevant in vivo. An assay was developed to determine the impact of antibody and cytotoxic cells on reducing the amount of measles virus cultured from infected cells. Although the components of this assay are the same as those involved in ADCC, the endpoint is a reduction in virus infectivity rather than cytotoxicity. The immune function measured in the assay has therefore been termed antibody-dependent cell-mediated immunity (ADCMI). Measles virus-infected Raji cells and blood mononuclear cells served as target and effector cells, respectively. Effector cells were incubated with antibody-labeled or unlabeled target cells for 24 h, and virus infectivity determined. Adding effector cells to unlabeled target cells reduced virus titer by 81.8%. Labeling target cells with measles-seronegative serum had little further effect. However, labeling target cells with measles-seropositive serum reduced infectivity an additional 96.5%. By allowing serum to remain in the supernatant fluid after labeling target cells, neutralizing and cell-mediated antibody functions were simultaneously measured. Finally, arming cytokine-activated effector cells with measles-seropositive serum also reduced virus infectivity. This novel assay provides an important tool for evaluating the anti-viral effects mediated by antibody and effector cells.

Lymphokine-activated cytotoxicity in peripheral blood mononuclear cells of severely immunocompromised HIV-infected patients.

The authors determined the natural killer (NK) and lymphokine-activated killer (LAK) activities of peripheral blood mononuclear cells (PBMCs) from a severely immunocompromised (CD4 cell counts < 100/mm3) group of AIDS patients, using K562 and U937 target cells. An increase in cytotoxicity was observed in the PBMCs of all 17 patients following a 48 h incubation with the combination of 400 U/ml of recombinant gamma interferon plus 20 U/ml of natural interleukin-2. Although NK and LAK activities were significantly higher in healthy controls than in patients, patients' LAK activity was higher than the NK activity of controls. The authors also demonstrated that the use of medium containing fetal bovine serum, when compared with medium containing autologous serum, increases NK activity without affecting LAK activity. Lymphokine augmentation of cytotoxicity is achievable in severely immunocompromised AIDS patients and might be of benefit in delaying opportunistic infections and malignancies.

Predominance of detrimental humoral immune responses to HIV-1 in AIDS patients with CD4 lymphocyte counts less than 400/mm3.

The humoral immune response to human immunodeficiency virus type 1 (HIV-1) was studied in 25 AIDS patients with CD4 lymphocyte counts of less than 400/mm3. Humoral immune responses against tissue culture adapted strains of HIV-1, and two limited-passage patient isolates were investigated. Total anti-HIV antibody levels were not significantly different between different individuals. Neutralizing titres against HIVLA1 and HIVSF2 were 10- to 100-fold higher than against clinical isolates. The complement-mediated, antibody-dependent enhancement of HIV-1 infection titre was high (mean 1:14,000). Antibody-complement mediated cytotoxicity of both HIVLA1 and HIVSF2 was ineffective using human complement as a complement source. The antibody-dependent, cell-mediated cytotoxicity (ADCC) activity varied against the four isolates with tissue culture-adapted strains being more susceptible than clinical isolates. Finally, an ADCC effector cell function, natural killer or NK activity, was measured for all 25 patients, and NK activity of patients was decreased by nearly 75% compared to uninfected individuals. In summary, beneficial humoral immune responses are low in HIV-1 infected individuals with CD4 counts of less than 400/mm3 if the in vitro assay system is constructed to best mimic the in vivo situation. These results suggest that the lack of functional antibody responses to HIV may play an important role in viral pathogenesis.

Age, sex, and household exposure are associated with the acute measles-specific antibody-dependent cellular cytotoxicity antibody response.

To determine whether functional antibody responses correlate with factors associated with severe measles, measles-specific antibody-dependent cellular cytotoxicity (ADCC) and neutralizing antibodies were measured in 114 Filipino children with measles. Children > 24 months old were more likely to have ADCC antibody in acute sera than were those < or = 24 months (odds ratio = 3.6, 95% confidence interval = 1.7-7.8). This age-related difference in ADCC prevalence was most apparent between younger and older girls. Among children < or = 24 months, a higher prevalence of ADCC antibody was associated with male sex, absence of lymphopenia, and household exposure to measles. The presence of ADCC antibody was not associated with malnutrition or diarrhea. Neutralizing antibody titers were lower in children with lymphopenia but showed no relationship with the other variables. Thus, the ADCC antibody response is associated with some risk factors related to measles severity. Attenuation of this response may contribute to the severity of infection.

Functional activities of 20 human immunodeficiency virus type 1 (HIV-1)-specific human monoclonal antibodies.

Antibodies that are useful in the treatment of HIV infection should result in virus neutralization or lysis of infected cells but should not enhance infection. In this study, the potential clinical use of 20 HIV-1-specific human monoclonal antibodies (HuMAbs) was determined by measuring their enhancing (C-ADE) activities using HIVLAI as the target virus. Two HuMAbs mediated both C-ADE and ADCC, two exclusively neutralized, and five exclusively mediated ADCC. Ten HuMAbs demonstrated no activity in any of the three assays. Three antibodies that neutralized HIVLAI were tested against HIVSF2; all three also neutralized HIVSF2. Four of five HuMAbs mediating ADCC against HIVLAI that were also tested against HIVSF2 had ADCC activity against HIVSF2. These results demonstrate that many HuMAbs have unique functions, allowing the separation of potentially beneficial and harmful activities. Combinations of HuMAbs with ADCC and neutralizing functions may have therapeutic utility.

Tratamento da deficiência de ferro em doadores de sangue: estudo comparativo entre sulfato ferroso e ferro proteinsuccinilato / Treatment of iron deficiency conditions in blood donors: controlled study of irn sulphate versus iron protein succinylate

ferro proteinsuccinilato é um novo preparado de ferro para administraçåo oral. Em estudo comparativo com o sulfato ferroso, em 40 doadores de sangue com baixos níveis de ferro de depósito, o tratamento por 30 dias com o ferro proteinsuccinilato resultou em maior absorçåo do ferro em comparaçåo com a droga de referência. A concentraçåo sérica de feroo aumentou significantemente em comparaçåo com os valores basais somente nos pacientes que receberam o ferro proteinsuccinilato. A quantidade de ferro de depósito, avaliada pelos níveis séricos de ferritina, aumentaram significativamente em ambos os grupos de tratamento

Measles virus-specific functional antibody responses and viremia during acute measles.

Antibody titers measured in functional and immunofluorescent assays were compared with proportions of peripheral blood mononuclear cells infected with measles virus in 8 adults with measles. In addition, a syncytium inhibition assay (SIA) for measuring neutralizing antibody using low-passage virus was compared with a standard plaque neutralization test (PNT). Antibody-dependent cellular cytotoxicity (ADCC) antibody rose later but attained higher titer than neutralizing, antibody-dependent complement-mediated lysis, IgM, or IgG antibodies. When titer changes between specimens from each patient obtained on different days were compared, only ADCC (r = .81, P = .026) and IgM (r = .81, P = .027) antibodies correlated with reductions in viremia. SIA and PNT correlated well (r = .93, P < .001). ADCC may be an important defense against measles. The delay in ADCC antibody relative to other antibodies is unique among viruses studied. The SIA is a useful alternative to the PNT for measuring measles neutralizing antibody.

Comparison of measles virus-specific antibodies with antibody-dependent cellular cytotoxicity and neutralizing functions.

Measles virus-specific antibodies with antibody-dependent cellular cytotoxicity (ADCC) and neutralizing functions were compared in 25 healthy women. ADCC antibody was measured in a 12-h 51Cr release assay using peripheral blood mononuclear cells as effectors and Raji cells persistently infected with measles as targets. Neutralizing antibody was determined by the plaque neutralization test (PNT). ADCC and PNT titers correlated well (r = .80, P < .001). ADCC titers ranged from < 10 to > 10(6) and averaged 1.3 logs higher than PNT titers. Three sera with PNT titers of < 120, including 1 with a titer of < 8 (< 10(0.9)), had ADCC titers of > or = 10(2.5). Among subjects born in the United States, ADCC titers, but not PNT titers, correlated inversely with the year of birth, suggesting that ADCC antibody may be more indicative than neutralizing antibody of differences between naturally and vaccine-acquired immunity. These data suggest a possible role for ADCC in protection or recovery from measles.

Comparison between first and second generation tests to determine the frequency of anti-HCV antibodies in uremic patients in replacement dialytic therapy.

Research was carried out on 74 hemodialysis patients to determine the frequency of anti-HCV antibodies; this was done by means of the 1st and 2nd generation screening and control tests. The 1st generation tests showed 9 HCV-seropositive patients, while the 2nd generation tests demonstrated 15 HCV-seropositive patients. It seems evident that the 2nd generation tests are more sensitive than those of the 1st.

Treatment of iron deficiency conditions in blood donors: controlled study of iron sulphate versus iron protein succinylate.

Iron protein succinylate is a new iron preparation for oral administration. In a controlled study versus iron sulphate in 40 blood donors with low levels of stored iron, treatment for 30 days with iron protein succinylate resulted in greater iron absorption compared to the reference drug. Serum iron concentration significantly increased compared with baseline values only in patients given iron protein succinylate. The amount of stored iron, evaluated by serum ferritin levels, significantly increased in both treatment groups.

Serum iron concentrations following administration of two different iron preparations.

Serum iron curves were determined in two groups of iron deficient patients after oral administration of iron protein succinylate or ferritin. The two preparations induced a significant increase of serum iron from 30 min after administration of a dose corresponding to 80 mg Fe3+. The increase induced by iron protein succinylate was more prolonged than that of ferritin.