Virologic and immunologic outcome of treatment of HIV infection with a herbal concoction, A-Zam, among clients seeking herbal remedy in Nigeria

Abstract: This study was to determine the effectiveness (CD4 count and viral load) of a safe herbal concoction, α-Zam used by clients seeking herbal remedy for treatment of HIV infection in Nigeria. 51 patients taking α-Zam as complementary and alternative therapy through the herbal therapist were studied for a period of 16 months. Preliminary medical and laboratory examinations using WHO and CDC criteria were done after confirmation of HIV infection by Western blotting in the nearest teaching hospitals to the residence of the patients. Regular visits were paid to the patients after commencement of the α-Zam to assess the side-effects, drug interactions, toxicity and effectiveness of the herbal remedy. There was a statistical significance (P<0.05) between pre-treatment and post-treatment CD4 count. 4 (7.8%) of the patients had average increase in CD4 count of 262±16 cell/µL, 23 (45.1%) patients with average increase 310±16 cell/µL, 16 (31.4%) patients with average increase 456±25 cell/µL and 8 (15.7%) patients with average increase 510±36 cell/µL( %) were at WHO staging I , II, III and IV respectively within 4 months on herbal therapy. There was very marked reduction in viral (HIV-RNA) load with 41 (80.4%) and 10 (19.6%) HIV infected patients had undetectable viral load and <1000 copies/ml respectively after the therapy. All symptoms and signs associated with HIV infection in all patients fully subsided within 4 weeks of commencement of α-zam therapy and there was no evidence of negative drug interaction in those HIV patients using both the herbal and highly active anti-retroviral therapy (HAART). The study is in progress to determine periodic immunological outcomes of post therapy in all patients

Hepatitis C virus prevalence and serotypes associated with HIV in The Gambia.

Hepatitis C virus (HCV) serotypes are important in the epidemiology and pathogenesis of HCV-related disease, but little is known of this connection in West Africa. Coinfection with human immunodeficiency virus (HIV) is associated with significant morbidity and mortality. This study aims to determine the prevalence of HCV and its serotypes associated with HIV in The Gambia. A total of 1500 individuals referred to the Royal Victoria Teaching Hospital for HIV serology between July and December, 2002 were screened for antibodies to HIV and subsequently for HCV, and seropositive samples were typed. This study shows HIV and HCV prevalence of 6.7% and 1.6%, respectively, with a co-infection rate of 0.6%. Serotype 2 showed the highest prevalence (58.1%), followed by serotype 1 (19.4%). Prevalence of HCV serotype 3 was 6.5% and five samples were untypeable. Co-infection of HIV-1 with HCV serotype 1 showed a prevalence of 44.4%, and with HCV serotype 2 of 33.3%. The findings support the evidence to suggest the West African subregion as the origin of HCV serotype 2. It also demonstrates the need for routine HCV screening of HIV-infected persons and blood donations, and calls for further studies to elucidate the sources of the HCV virus.

Influence of tart cherry juice on indices of recovery following marathon running.

This investigation determined the efficacy of a tart cherry juice in aiding recovery and reducing muscle damage, inflammation and oxidative stress. Twenty recreational Marathon runners assigned to either consumed cherry juice or placebo for 5 days before, the day of and for 48 h following a Marathon run. Markers of muscle damage (creatine kinase, lactate dehydrogenase, muscle soreness and isometric strength), inflammation [interleukin-6 (IL-6), C-reactive protein (CRP) and uric acid], total antioxidant status (TAS) and oxidative stress [thiobarbituric acid reactive species (TBARS) and protein carbonyls] were examined before and following the race. Isometric strength recovered significantly faster (P=0.024) in the cherry juice group. No other damage indices were significantly different. Inflammation was reduced in the cherry juice group (IL-6, P<0.001; CRP, P<0.01; uric acid, P<0.05). TAS was ~10% greater in the cherry juice than the placebo group for all post-supplementation measures (P<0.05). Protein carbonyls was not different; however, TBARS was lower in the cherry juice than the placebo at 48 h (P<0.05). The cherry juice appears to provide a viable means to aid recovery following strenuous exercise by increasing total antioxidative capacity, reducing inflammation, lipid peroxidation and so aiding in the recovery of muscle function.

Prevalence of HIV-1, HIV-2, hepatitis C and co-infection in The Gambia.

BACKGROUND: In most West African countries, the distribution and risk factors for co-infection with Human Immunodeficiency Virus (HIV) and Hepatitis C Virus (HCV) is unknown despite the current HIV epidemic and evidence of increasing prevalence of HCV in the region. OBJECTIVE: This study aimed to evaluate the distribution and the risk factors for the transmission of co-infection between HIV and HCV in The Gambia. METHODS: A total of 1500 persons referred for HIV serology at the Royal Victoria teaching Hospital were interviewed following informed consent to obtain information on their demographic variables, knowledge of sexually transmitted diseases and their prevention, and patterns of risk behavior. Blood was collected and tested for anti-HIV and anti-HCV antibodies by Enzyme Linked Immunosorbent Assay (ELISA). RESULTS: In the general population, the prevalence of HIV was 6.7%, while that of HCV was 2.1%. Both infections occurred more frequently in males than in females. HIV and HCV coinfection rate was 0.6%. Co-infection was significantly more common in males than females. All types of infection--HIV, HCV and HIV/HCV co-infections occurred much more in polygamous settings than in monogamy. CONCLUSION: This study has demonstrated the extent of coinfection with HIV and HCV in The Gambia. The prevalence of female circumcision may be a contributory occurrence factor in the transmission of HIV but not in that of HCV.

CD4+ lymphocyte values and trends in individuals infected with human immunodeficiency virus and/or co-infected with hepatitis C virus in the Gambia.

OBJECTIVES: This study was undertaken to monitor the CD4+ lymphocyte count in individuals infected with Human Immunodeficiency Virus (HIV) and/or co-infected with Hepatitis C Virus (HCV) and to compare this with the counts in normal individuals in The Gambia. METHODS: Blood samples were taken from 1500 individuals referred for HIV serology at the Royal Victoria Teaching Hospital (RVTH) following informed consent. Samples were tested for antibodies to HIV by the Murex ELISA, antibodies to HCV by the Ortho ELISA, and CD4 counts determined by the Dynalimmunomagnetic cell isolation method RESULTS: Of the 1500 patients screened for HIV and HCV antibodies, 6.7% (101/1500) were infected with HIV, 0.6 % (9/1500) were co-infected with HCV and 1.5 % (22/1500) were infected with HCV alone. Almost half (44.6%; 25/56) of HIV-1 infected patients had a CD4+ lymphocyte count at diagnosis of 200 cells/microl or less as compared to 41.7 % (10/24) of HIV-2 and 75% (6/8) of HIV-D infected patients. The rate of CD4 decline was higher among HIV/HCV co-infected persons than individuals infected with HIV or HCV. The rate of decline was higher among men than women. These differences did not reach statistical significance due in large part to the small number of participants who completed the programme. The CD4+ lymphocyte count of apparently healthy Gambian male and females was 489 cells/microl and 496 cells/microl respectively. This rate is lower than that reported for Caucasians, but in agreement with the global range. CONCLUSION: A significant progressive decline in CD4+ lymphocyte count was observed among the female control group who were negative for HIV and HCV. This finding is unclear and calls for a longitudinal study involving a cohort of women in this region.

Induction of oxidative DNA damage by Helicobacter pylori in HT29 cells.

Infection with Helicobacter pylori has been associated with the development of gastric adenocarcinoma in humans. Several routes have been implicated, the main one being oxidative DNA damage resulting from chronic inflammation, which accompanies infection. However, DNA has been demonstrated in human cells after in vitro incubation with H. pylori sonicates. Using the fragment length analysis using restriction enzymes (FLARE) assay, this study investigates the DNA damaging potential of three clinical isolates of H. pylori on cultured HT29 cells. Significant amounts of oxidative DNA damage were detected in HT29 cells following a 72-hour incubation with each H. pylori isolate. As tumour induction is a known consequence of oxidative DNA damage, chronic infection with the organism may lead to the development of adenocarcinoma of the stomach.

Preferential survival of acute lymphoblastic leukemia cells at 33 degrees C is associated with up-regulation of bcl-2.

An important feature of childhood acute lymphoblastic leukemia (ALL) is the risk of testicular relapse in affected males, which may occur months or years after induction of remission. However, little is known about the factors that regulate leukemic cell survival and resistance to chemotherapy in the testis. In the present study, incubating ALL cell lines and primary cells from ALL patients at 33 degrees C resulted in increased survival, resistance to chemotherapeutic agents and upregulation of bcl-2. Acute myeloid leukemia cell lines incubated at 33 degrees C also showed increased survival and resistance to chemotherapeutic agents, but did not demonstrate upregulation of bcl-2. This may be important in determining survival of ALL cells at lower temperatures in the testis.

Human immunodeficiency virus and hepatitis C co-infection in sub-Saharan West Africa.

Co-infection with human immunodeficiency virus (HIV) and hepatitis C virus (HCV) is becoming a major global problem, leading to increased morbidity and mortality in developed countries. Co-existence in sub-Saharan West Africa of a high prevalence of HIV and HCV, which share similar behavioural risk factors and modes of transmission, must be seen in the broader context of an emerging third epidemic of HIV and HCV co-infection, as many factors that may affect the spread of HIV and HCV co-infection are endemic in the continent, including host factors such as sexual behaviour, presence of other sexually transmitted diseases, female and male circumcision status, percutaneous and perinatal exposure, and poverty. This review examines the epidemiology, risk factors and transmission of HIV and HCV co-infection and draws attention to the possible emergence of an epidemic of HIV and HCV co-infection in the region.

Human breast milk immunology: a review.

Breast feeding has been shown to enhance the development of the immune system of the newborn as well as provide protection against enteric and respiratory infections. It has been suggested that implementation of breast feeding programs has the potential to save hundreds of thousands of lives worldwide. Human milk is a bodily fluid which, apart from being an excellent nutritional source for the growing infant, also contains a variety of immune components such as antibodies, growth factors, cytokines, antimicrobial compounds, and specific immune cells. These help to support the immature immune system of the newborn baby, and protect it against infectious risks during the postnatal period while its own immune system matures. This article reviews some of the factors in human breast milk that give it these important properties.

Differential expression of CD 180 and IgM by B-cell chronic lymphocytic leukaemia cells using mutated and unmutated immunoglobulin VH genes.

We have studied the surface expression of the Toll-like receptor family member CD 180 on cells from 78 patients with B-chronic lymphocytic leukaemia (B-CLL). B-CLL cells had variable levels of CD 180 expression, but this was always less than that expressed by normal blood B cells and was stable for 24 months. Significantly higher levels of CD 180 were expressed by B-CLL cells with mutated IGVH genes compared with those using unmutated IGVH genes. This was in contrast to the higher levels of expression of surface immunoglobulin M by B-CLL cells using unmutated, rather than mutated IGVH genes. CD 180 was functional on B-CLL cells from some of the patients, as shown by the increased expression of CD 86 following incubation in vitro with anti-CD 180. The differential expression of CD 180 amongst B-CLL patients is one more marker that may define more precisely the different biological properties of this heterogeneous disease.

Hepatitis C antibodies in asymptomatic first-time blood donors in The Gambia: prevalence and risk factors.

This study evaluates the seroprevalence and risk factors for hepatitis C (HCV) antibodies in asymptomatic first-time blood donors in The Gambia. The study population includes 460 blood donors (age range: 18-40 years [mean: 27.5]) who attended the Royal Victoria Teaching Hospital from July to December 2002. Antibodies to hepatitis C are determined using and enzyme-linked immunosorbent assay (ELISA) test system. The prevalence of hepatitis C found in this study was 1.1% (95% CI, 0.16-1.12). Previous history of sexually transmitted disease, married men in polygamous relationships, and hospital or clinic-based workers were determined to be at risk of acquiring hepatitis C. The study shows that seroprevalence of hepatitis C in The Gambia is low compared to other countries in the region.

Is the liver an important site for the development of immune tolerance to tumours?

Despite the enormous interest in the field of tumour immunology, and the development of vaccine based strategies for immunotherapy of tumours, results in patients with cancer have been disappointing. This is partly due to the lack of development of clearly defined anti-tumour immune responses. The basis for the induction of specific anti-tumour non-responsiveness is not known. Recently, the liver has been recognised as an important organ in the regulation of peripheral immunological responses. It is characterised by a remarkable ability to induce tolerance to antigens from a variety of sources. Oral tolerance to food antigens, antigens from gut flora and other antigens administered via the oral route is partly dependent upon local immunoregulation in the liver. Transplantation of liver tissue shows a remarkable ability to induce tolerance in some species, not only to liver tissue but also to other organs and tissues transplanted at the same time. This tolerance can be transferred by adoptive transfer of lymphocytes. It has been suggested that the establishment of persistent infection in the liver by hepatitis viruses, may partly depend on the tolerogenic environment of the liver, and that this may also play a role in the development of hepatocellular carcinoma in patients with chronic infections with these viruses. The liver is also a common and an important site for the development of metastases from many primary tumours. This is partly dependent upon the anatomic location and structure of the liver, but may also partly reflect the exploitation of the tolerogenic environment in the liver, allowing micrometastases to colonise and grow. This may account for the fact that the liver is such a common site for metastasis. Furthermore, once tolerance to tumour antigens is established in the liver, tolerated lymphocytes may migrate from the liver back to primary tumours and exacerbate immunological non-responsiveness at tumour sites. Indeed, if this happens early in tumour development, liver dependent tolerance to tumour antigens may play a significant role in tumour progression, and may partly determine impaired tumour responses in vaccine based immunotherapy strategies.

Leucodepletion for transmissible spongiform encephalopathies.

Transmissible spongiform encephalopathies (TSEs) have been recognised around the world for many years. Creutzfeldt-Jakob disease (CJD), one of the human forms of TSE, has been studied widely and thus far has not proved a great threat to human health. The emergence of two new TSEs--bovine spongiform encephalopathy (BSE) in cattle and variant Creutzfeldt-Jakob disease (vCJD) in humans in the UK--has caused great concern. BSE has had an economic impact and vCJD is a threat to human health. It has been shown that these two diseases are caused by the same prion agent and are linked. Research indicates that vCJD behaves differently to CJD and there is strong evidence to suggest that vCJD is present in lymphoid tissues and B lymphocytes, which presents a theoretical risk that it may be transmitted by transfusion of blood and blood products. To minimise/prevent this risk, the UK government has decided that plasma should be sourced from abroad and has instructed the National Blood Service to leucodeplete all blood and blood products, at a cost of 70 million pounds per annum, although it is not known if this will remove this risk.

Transendothelial migration confers a survival advantage to activated T lymphocytes: role of LFA-1/ICAM-1 interactions.

The clearance of activated T lymphocytes by apoptosis is an essential component in the resolution of the immune response; however, certain signals received within inflamed tissue may result in the persistence of activated T cells. Our previous work has shown that, when compared with resting cells, effector cells migrate more efficiently across endothelium, thus such cells may be selectively recruited to sites of inflammation. We hypothesized that transmigration of T cells across endothelium might influence cell survival. We have generated T cell lines by culturing in IL-2 following PHA activation. These T cell lines die rapidly by apoptosis when deprived of IL-2 (53.7 +/- 4.0% survival after 24 h). In contrast, cells that have migrated across human umbilical vein endothelial cells (HUVEC) survived significantly better than control cells (80.3 +/- 3.6%, n= 18, P<0.001). Endothelial cell conditioned medium was also able to reduce apoptosis, but this effect was small when compared with the protective effect of transmigration. Culture of T lymphocytes on fibronectin, or RGD peptides, or in suspension with a range of chemokines active on T cells, including RANTES and lymphotactin had no effect on survival. In contrast, blocking LFA-l/ICAM-l interactions reduced the protective effect of transmigration (42.3 +/- 6.7% reduction). Culture of activated T cells on immobilized ICAM-l alone also increased survival. These results indicate that signals received by activated T cells during extravasation can influence their subsequent survival within tissue, and implicates the involvement of LF A-l/ICAM-l interactions.

Expansion of CD4+ T cells with a cytotoxic phenotype in patients with B-chronic lymphocytic leukaemia (B-CLL).

Abnormal CD4/CD8 ratios and T-cell function have previously been shown in patients with B-chronic lymphocytic leukaemia (B-CLL). We have demonstrated that CD4+ T cells containing both serine esterase and perforin (PF) are increased in the blood of these patients. Using flow cytometry, we have shown that the CD4+ PF+ cells were CD57+ but lacked expression of CD28, suggesting a mature population. The same phenotype in CD8+ T cells is characteristic of mature cytotoxic T cells. However, in contrast to the CD8+ T cells, the CD4+ T cells were more frequently CD45RO positive than CD45RA positive, indicating prior antigen experience. In contrast, this population lacked expression of either CD69 or HLA-DR, arguing that they were not activated or that they are an abnormal population of T cells. Their constitutive cytokine levels showed them mainly to contain IL4 and not IFNgamma, suggesting a Th2 phenotype. The role of the CD4+ PF+ T-cell population is at present uncertain. However, this potentially cytotoxic T-cell population could contribute both to enhancing survival of the B-CLL tumour cells through production of IL4, and to the immunodeficient state frequently seen in patients with this tumour, independent of drug treatment.

CD5 B cells and B-cell malignancies.

Over the past year, progress has been made in understanding of the physiology and disease associations of CD5+ (B1) B cells, although their exact role in pathogenesis remains unclear. Earlier studies on the negative function of CD5 within the B-cell receptor complex have been substantiated, and it seems likely that soon the signaling pathways used by this coreceptor will be elucidated. Progress in diagnosis, physiology, and etiopathogenesis of CD5+ malignancies has been made, particularly in B-cell chronic lymphocytic leukemia. The low-level expression of surface immunoglobulin has been explained by the mutations that occur in the associated CD79b. Two new potential tumor-suppressor genes have been identified in the hot spot of chromosome 13q, which provides an exciting step forward in understanding of the etiopathogenesis of some B-cell chronic lymphocytic leukemia. Activated signal transducers for activation of transcription factors molecules have been shown to be phosphorylated on different amino acids in B1 and chronic lymphocytic leukemia tumors, although the significance of this is, as yet, unclear. Finally, aberrant expression of CD40L by chronic lymphocytic leukemia T cells may contribute to the immunodeficiency that develops in these patients.

Serum levels of TNF, IL-6 and sCD23 correlate with changes in lymphocyte count in patients with B-cell chronic lymphocytic leukaemia receiving interferon-alpha therapy.

The control of cell growth and differentiation in B-cell malignancies may be regulated by the autocrine production of cytokines, several of which have been implicated in the growth and survival of B-cells. The effect of interferon-alpha (IFN) therapy in these disorders may be to disrupt autocrine growth or survival loops. We have measured levels of circulating IL-1b, IL-6, TNF-a and soluble CD23 (sCD23) in 8 patients with Binet stage A B-cell chronic lymphocytic leukaemia (B-CLL) receiving IFN therapy, and compared these with changes in the lymphocyte count following IFN therapy. Two patients developed anti-interferon antibodies while on IFN therapy, and in both them, the changes in lymphocyte count correlated significantly with the titre of anti-interferon antibodies, as well as serum levels of IL-6, TNF-a and sCD23. In one patient there was significant correlation with levels of IL-1b. One patient, who stopped and restarted IFN therapy, demonstrated correlation between lymphocyte count and levels of IL-6 and sCD23. In a further two patients, there was correlation with levels of sCD23 alone, while the remaining three patients showed no correlation between lymphocyte count and the serum cytokines measured. These results suggest that IFN therapy may alter levels of circulating cytokines in some CLL patients and that these effects may be associated with disease progression.

Regulation and function of adhesion molecules in B-cell chronic lymphocytic leukaemia.

Cell surface expression of adhesion molecules in B-cell chronic lymphocytic leukemia (B-CLL) may determine the patterns of dissemination and infiltration. B-CLL cells express high levels of CD44, but expression of the leucocyte integrins was low or absent, while expression of VLA-4 is high. Most cases examined expressed no detectable ICAM-1, but some cases demonstrated levels of up to 30%. Levels of L-selectin were also variable, and expression could be induced/enhanced in vitro by incubation with cytokines such as IL-4, interferon-alpha and interferon-gamma. B-CLL cells bound normally to vascular endothelium, but binding to IL-1-activated endothelium was significantly lower than that of normal peripheral blood lymphocytes. Cytokine enhancement of L-selectin expression was not accomplished by changes in binding to vascular endothelium. Patterns of adhesion molecule expression and their regulation by cytokines may underly some of the clinical features of this disease.

Cutaneous manifestation of leishmaniasis 40 years after exposure in a patient with chronic lymphocytic leukaemia.

Most individuals infected with the protozoan parasite Leishmania do not demonstrate overt disease, because of effective immune protection produced by T-cell mediated immunity. Acquired defects in T-cell responses may lead to emergence of leishmaniasis many years after exposure in endemic areas. We describe a case of a 75 year old man, who presented with a cutaneous manifestation of leishmaniasis 40 years after exposure, co-incident with the diagnosis of chronic lymphocytic leukaemia.

Interferon-alpha, Bcl-2 expression and apoptosis in B-cell chronic lymphocytic leukemia.

Interferon-alpha has been used as therapy in patients with B-cell chronic lymphocytic leukaemia (B-CLL), and is able to induce remissions in patients with early stage disease. Although interferon-alpha exhibits a wide variety of cellular effects, none of these have adequately explained the mechanism of action of interferon-alpha in B-CLL. Recent attention has focussed on the role of bcl-2 in B-CLL, and the regulation of bcl-2 expression by cytokines. B-CLL is characterized by the relentless accumulation in the peripheral blood and bone marrow of a monoclonal population of long-lived B-cells. However, when these cells are cultured in vitro, they die rapidly by apoptosis or programmed cell death. It has recently been demonstrated that B-CLL cells can be protected from apoptotic death in vitro by co-culture with cytokines, such as IL-1, IL-2, IL-4, IL-6 and interferon-gamma. The protection against apoptosis is correlated with increased levels of bcl-2 expression. It was suggested that interferon-alpha induced remissions in early stage B-CLL by interrupting these growth-factor dependent survival pathways and allowing the cells to die by apoptotic death in vivo. However, interferon-alpha has also been shown to protect B-CLL cells from apoptotic death in vitro. This suggests that interferon-alpha does not produce remission in B-CLL by direct effects on B-CLL cells in the circulation. Many of the cytokines which protect B-CLL cells from apoptotic cell death, are members of the cytokine receptor family which utilize a common family of signal transduction molecules. Further elucidation of these signal transduction pathways may offer the prospect of developing novel therapeutic strategies aimed at inducing apoptosis of the malignant clone in vivo.