Augmented Innate and Adaptive Immune Responses Under Conditions of Diabetes-Filariasis Comorbidity.

Metainflammation, as seen in chronic diabetes subjects, impairs immunity and increases the susceptibility to infections. In the present study, the effect of diabetes on immune response against filariasis was studied. Both toll-like receptor (TLR)-mediated and crude antigen-induced immune responses were quantified, in whole blood cultures from filariasis-infected subjects (LF+), with and without diabetes. Blood cultures were stimulated with TLR ligands (TLR2 and TLR4) or filarial antigen or were left unstimulated (control) for 18 h. Cytokine, chemokine, and defensin secretion was quantified by ELISA. Expression of HLA-DR, B7-1, B7-2, activation marker (CD69), and Th (Th1, Th2, Th17, and Th9) phenotypes was quantified by flow cytometry. Expression of immunomodulatory effectors (Cox-2, HO-1, IDO-1, and p47Phox) and Th-polarizing transcription factors (T-bet, GATA3, and ROR-γt) was quantified by quantitative PCR. Secretion of IL-27, IL-1Ra, IL-12, IL-33, IL-9, and SDF-1 was increased under diabetes conditions with increased Th9 polarization and increased expression of Cox-2 and IDO. Overall, diabetes was found to augment both TLR-mediated and antigen-induced inflammation, which can promote chronic pathology in LF+ subjects.

Highlighting the Relevance of CD8+ T Cells in Filarial Infections.

The T cell immune responses in filarial infections are primarily mediated by CD4+ T cells and type 2-associated cytokines. Emerging evidence indicates that CD8+ T cell responses are important for anti-filarial immunity, however, could be suppressed in co-infections. This review summarizes what we know so far about the activities of CD8+ T cell responses in filarial infections, co-infections, and the associations with the development of filarial pathologies.

Inflammatory responses induced by the filarial nematode Brugia malayi are mediated by lipopolysaccharide-like activity from endosymbiotic Wolbachia bacteria.

The pathogenesis of filarial disease is characterized by acute and chronic inflammation. Inflammatory responses are thought to be generated by either the parasite, the immune response, or opportunistic infection. We show that soluble extracts of the human filarial parasite Brugia malayi can induce potent inflammatory responses, including tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta, and nitric oxide (NO) from macrophages. The active component is heat stable, reacts positively in the Limulus amebocyte lysate assay, and can be inhibited by polymyxin B. TNF-alpha, IL-1beta, and NO responses were not induced in macrophages from lipopolysaccharide (LPS)-nonresponsive C3H/HeJ mice. The production of TNF-alpha after chemotherapy of microfilariae was also only detected in LPS-responsive C3H/HeN mice, suggesting that signaling through the Toll-like receptor 4 (TLR4) is necessary for these responses. We also show that CD14 is required for optimal TNF-alpha responses at low concentrations. Together, these results suggest that extracts of B. malayi contain bacterial LPS. Extracts from the rodent filaria, Acanthocheilonema viteae, which is not infected with the endosymbiotic Wolbachia bacteria found in the majority of filarial parasites, failed to induce any inflammatory responses from macrophages, suggesting that the source of bacterial LPS in extracts of B. malayi is the Wolbachia endosymbiont. Wolbachia extracts derived from a mosquito cell line induced similar LPS-dependent TNF-alpha and NO responses from C3H/HeN macrophages, which were eliminated after tetracycline treatment of the bacteria. Thus, Wolbachia LPS may be one of the major mediators of inflammatory pathogenesis in filarial nematode disease.

A new insight into the pathogenesis of filarial disease.

Filariasis is a major public health problem throughout many regions of the tropics. The disease is caused by several species of filarial nematode including Wuchereria bancrofti and Brugia malayi, the agents of lymphatic filariasis, and Onchocerca volvulus, the cause of 'riverblindness'. Disease caused by these worms varies depending on the tissue location of the parasite, and is associated with episodes of acute and chronic inflammation. These pathologies, including elephantiasis and blindness, rank among the most disabling in the world. Studies aimed at characterizing the molecular nature of the inflammatory stimuli derived from filarial nematodes uncovered a long forgotten secret, their symbiont Wolbachia. LPS-like molecules from these intracellular bacteria are responsible for potent inflammatory responses from macrophages and in animal models of filarial disease. Wolbachia has also been associated with severe inflammatory reactions to filarial chemotherapy, being released into the blood following the death of the parasite. Recent studies in animal models even implicate Wolbachia in the onset of lymphodema and blindness. Taken together these studies suggest a major role for Wolbachia in the pathogenesis of filarial disease. It may be possible, through the use of antibiotic therapy, to clear worms of their bacteria, in the hope that this will prevent the onset and development of filarial pathology.

Decline of brugian filariasis in Cherthala taluk, Alappuzha district, Kerala.

A total of 4492 persons from 5 panchayats and 1 town were investigated from the Brugia malayi most endemic taluk of Cherthala, Alappuzha district of Kerala state. The urban area in Cherthala taluk only revealed mf carriers; mf rate was 0.13%. Rural areas in Cherthala taluk were free from infection. Microfilaria rate had declined by 99.5% and disease rate by 90.7% in Cherthala compared to 1934 prevalence. Shedding of sheath by B. malayi microfilariae was recorded for the first time in India. The youngest person with microfilaria and disease manifestation was 4 1/2 and 9 years respectively. All the 3 major vectors, Mansonia annulifera, Ma.uniformis and Culex quinquefasciatus were prevalent throughout. Complete disappearance of brugian filariasis from this taluk is a distinct possibility. The reasons for the drastic decline are discussed.

Maternal Infection Is a Risk Factor for Early Childhood Infection in Filariasis.

BACKGROUND: Global Program to Eliminate Lymphatic Filariasis (GPELF) launched by WHO aims to eliminate the disease by 2020. To achieve the goal annual mass drug administration (MDA) with diethylcarbamazine (DEC) plus albendazole (ABZ) has been introduced in all endemic countries. The current policy however excludes pregnant mothers and children below two years of age from MDA. Since pregnancy and early childhood are critical periods in determining the disease outcome in older age, the present study was undertaken to find out the influence of maternal filarial infection at the time of pregnancy on the susceptibility outcome of children born in a community after implementation of MDA for the first time. METHODOLOGY AND PRINCIPAL FINDINGS: The participants in this cohort consists of pregnant mothers and their subsequently born children living in eight adjacent villages endemic for filarial infections, in Khurda District, Odisha, India, where MDA has reduced microfilariae (Mf) rate from 12% to 0.34%. Infection status of mother and their children were assessed by detection of Mf as well as circulating filarial antigen (CFA) assay. The present study reveals a high rate of acquiring filarial infection by the children born to infected mother than uninfected mothers even though Mf rate has come down to < 1% after implementation of ten rounds of MDA. SIGNIFICANCE: To attain the target of eliminating lymphatic filariasis the current MDA programme should give emphasis on covering the women of child bearing age. Our study recommends incorporating supervised MDA to Adolescent Reproductive and Sexual Health Programme (ARSH) to make the adolescent girls free from infection by the time of pregnancy so as to achieve the goal.

Population dynamics of microfilarial production and eosinophilic levels in slow lorises infected with Breinlia sergenti. Petter (Filarioidea: Dipetalonematidae).

Population dynamics of microfilarial production and eosinophilic levels in slow lorises infected with Breinlia sergenti, Petter (Filarioidea: Dipetalonematidae). International Journal for Parsitology 4: 383388. Observations have been made on microfilarial and eosinophilic levels in slow lorises infected with Breinlia sergenti. Animals given a single inoculation of 100-150 infective larvae exhibited three different patterns of microfilaraemia while superinfected animals showed enhanced microfilarial levels. It appeared that the number of inoculations as well as the interval between inocula are important factors in enhancing microfilarial levels. Two different types of incubation periods were seen, one at 100-120 days and the other at 200 days. The eosinophilic levels were investigated in some of the animals and an attempt was made to correlate these levels with the microfilaraemia. Cortisone injection appeared to promote a vigorous eosinophilia in some of the infected animals tested.

Short report: total serum levels of the nitric oxide derivatives nitrite/nitrate during microfilarial clearance in human filarial disease.

Nitric oxide (NO) has recently been shown to be cytotoxic to both microfilariae and adult Brugia malayi in vitro and in a murine model, as well against Onchocerca lienalis microfilariae in vitro. We studied the kinetics of nitrite/nitrate, both stable end products of NO, by high-pressure liquid chromatography during microfilaricidal chemotherapy in four filariasis (three Loa loa, and one Onchocerca volvulus) patients. High serum levels of nitrite/nitrate were released during microfilarial clearance and sustained elevated levels were observed six months after chemotherapy, suggesting a role of NO in the elimination of microfilariae in human filariasis.

Longitudinal analysis of the development of filarial infection and antifilarial immunity in a cohort of Haitian children.

Longitudinal studies are being conducted in Leogane, Haiti to investigate the relationship between acquisition of filarial infection and development of antifilarial immunity as well as the impact of maternal infection on this relationship. Children (0-24 months of age) residing in Leogane were enrolled and were examined periodically to monitor parasitologic status and to collect serum for antigen and antifilarial antibody determinations. To examine the development of filarial antigenemia and antifilarial antibody responses in this cohort, serum samples were selected from a cross section of the population at two (n = 82) and four years of age (n = 76). Antigen prevalence increased from 6% among two-year-olds to more than 30% among four-year-olds, but in only one four-year-old child were microfilaria detected in a 20-microl smear. The proportion of antigen-positive children born to antigen-positive mothers was higher than the proportion of antigen-positive children born to antigen-negative mothers (9.8% versus 0% for two-year-olds; P = 0.15; and 39.6% versus 22.7% for four-year-olds; P = 0.18). Antifilarial IgG4 levels were significantly higher among antigen-positive children at both two and four years of age (P < 0.001). In analyses of paired samples, antifilarial IgG4 responses increased significantly more among children who acquired infection by four years of age than among children who remained antigen negative, whereas antifilarial IgG1 and IgG2 responses changed equally for antigen-positive and -negative children. Antifilarial antibody levels were not influenced by maternal infection status, but were significantly influenced by age, antigen status, and the neighborhood within the community. These results provide evidence that children acquire infection early in life and suggest that antifilarial antibody responses may peak in early childhood.

Filarial elephantiasis in French Polynesia: a study concerning the beliefs of 127 patients about the origin of their disease.

127 patients from Tahiti who were suffering from elephantiasis were interviewed about their opinion of the origin of their disease. Ancestral beliefs are still widely held even after 25 years of antifilarial campaigns which have resulted in a drastic decrease in endemicity with almost no clinical incidence. It is disappointing that the responsibility of mosquitoes is denied by a majority of patients. The explanations are to be found in the unusual evolution of this disease and in the small importance attached to sanitary education.

Effect of pyridoxine deficiency on Litomosoides carinii infection in albino rats.

Pyridoxine deficiency has been induced in albino rats at the weanling or neonatal stages. Exposure of the vitamin-deficient rats to the infective larvae of Litomosoides carinii resulted in no development or establishment of the parasite in the host. Although the food consumption of the pair-fed group was necessarily restricted to that of the deficient group, the susceptibility of the animals of the pair-fed group receiving the normal vitamin intake resembled that of the control group. The humoral immune response to the infection in the pair-fed controls as measured by indirect haemagglutination, was inhibited compared to that in the controls. This inhibition was more severe in the vitamin-deficient animals.

The impact of housing structures on filarial infection.

A study was undertaken to correlate the impact of housing and patterns of house construction on the vector density and transmission of filaria among the inhabitants of these houses. Three different types of houses in ecologically similar hamlets of Hariharpur village in Varanasi were selected for determining the density of Culex quinquefasciatus, the vector of Wuchereria bancrofti and its infectivity. The maximum per man hour density of the vector was recorded during March (31.66, 40.33 and 41.33) while minimum was recorded during June (1.3, 2.6 and 0.33) in all the three types of houses. Infection rate in the vectors collected from poorly constructed houses was observed during April, May, October and January of the following year, whereas in moderately constructed houses, infection was observed only in September and in the well constructed houses dissection results did not reveal any infection during the months of the study. Infectivity rate was observed to be 10.0% in moderately constructed houses (group B) during the month of September and 14.2% in poorly constructed houses (group C) during the month of October. Parasitological observations of the population showed a 12.2% microfilaria (mf) rate and 6.7% disease rate among the residents of poorly constructed houses, 5.8% mf rate and 2.9% disease rate among residents of moderately constructed houses. Among residents of well built houses (Group A), none were found to be positive with mf, but disease rate was observed to be 2.7%. Throughout the year the relative humidity was observed to be higher in the poorly constructed houses and ambient temperatures were found to be lower during the summer but higher during the winter than to those of the better constructed houses. The study made evident that the construction of houses plays an important role in the vector's resting preference, leading to a higher density in poorly constructed houses, thereby increasing the possibility of infection within them, and thus maintaining a higher potential for filarial transmission among its inhabitants.

Edema resulting from experimental filariasis.

Domestic cats and patas monkeys were infected with Brugia malayi so that the worms localized in the regional lymphatics of the hind legs. Reaction to the filarial parasites resulted in visible local edema in cats and disruption of normal lymph flow in the monkeys. Edematous tissue was examined grossly and by light and electron microscopy. Lymph flow patterns were examined by direct observation following injection of lymph staining dye and reflection of the skin, by X-ray following injection of radio-opaque contrast media, and by lymphscintigraphy after subcutaneous injection of radioisotopes. Clinical edema occurred in cats but not in monkeys. However, disruption of normal lymph flow in monkeys infected with Brugia could be demonstrated by lymphscintigraphy.

Filaroides hirthi in a dog: fatal hyperinfection suggestive of autoinfection.

Progressive dyspnea in a dog began 1 month after it had been hit by a car. Radiographs revealed diffuse interstitial and alveolar pneumonia. Large numbers of embryonated eggs were observed in bronchial washings; and many small nematodes, later identified as Filaroides hirthi, were found in the fine needle aspirates of pulmonary tissue. Numerous nematode larvae were demonstrated on fecal flotation. The severe hyperinfection strongly suggested autoinfection, probably associated with depressed immunologic competence due to chronic stress.

Parafilariosis in African buffaloes (Syncerus caffer).

This is the first report on the occurrence of Parafilaria bassoni in the African buffalo (Syncerus caffer). Previously this parasite has been recorded only in springbok (Antidorcas marsupialis) in Namibia. Haemorrhagic perforations (bleeding points), the usual lesions seen in infected animals, were caused by gravid female parasites ovipositing embryonated eggs. These lesions occurred mainly on the dorsal and lateral sides of buffaloes. Complications of these lesions developed in a small number of buffaloes because of secondary bacterial infection [subcutaneous abscesses (3/178)] and as a consequence of a localized Type 1 hypersensitivity [large cutaneous ulcers (7/178)]. Red-billed oxpeckers (Buphagus erythrorynchus) appeared to play an important role in the epidemiology of this parasite as well as in the pathogenesis of the lesions. They reduced the likelihood of spread by ingesting blood containing embryonated eggs, and caused the development of large ulcers by feeding on superficial necrotic skin. From the results of an ELISA test it was determined that P. bassoni-infected buffaloes occur throughout the Kruger National Park complex, with a seroprevalence of approximately 34%.

Brugia timori: experimental infection in some laboratory animals.

Experimental infection with Brugia timori of 7 jirds (Merionesunguiculatus), 4 cats and 2 monkeys (Macaca fascicularis) is described. Although no microfilariae were detected by examining 20 microliter samples of tail blood of jirds, adult worms were recovered from 6 of the 7 jirds at autopsy 69-141 days following infection. Some worms were gravid and microfilariae were found in visceral blood of 2 animals. The adult recovery rate in jirds was 16%; the male to female ratio was 1:3. In cats patent infection developed in 95-105 days but microfilaraemias were of low level and transient. No parasites were recovered from monkeys.

Experimental Brugia malayi infections in the rhesus monkey.

Twenty-eight rhesus monkeys in 3 groups were exposed to single (Group I), double (Group II), and multiple (Group III) inoculations with B. malayi infective larvae. Infections were monitored by microfilarial and blood counts, selected biochemical tests, IFA responses, and records of body temperature and lymphadenopathy before and/or after treatment with DEC. As a whole, the highest microfilaraemia levels were observed in Group II and lowest in Group III monkeys. Eosinophilia was a common occurrence but reached the highest mean levels in Group III. Intermittent fevers and lymph node enlargements were observed in all groups of monkeys and the occurrence of these appeared to be correlated. No definite pattern of antibody production was discernable among groups, but an inverse relationship existed between microfilaraemia and detectable microfilarial antibodies. Treatment with DEC produced a microfilaraemia-taxic effect within the initial half hour and responses to treatment varied according to individuals. Although post-treatment reinfection appeared to cause lymphoid responses and tissue eosinophilia, no substantial resistance to reinfection was observed.

[Bacterial symbionts (Wolbachia) of filarial nematodes: implications for the treatment and pathology of filariasis]. / I simbionti batterici (Wolbachia) delle filarie: implicazioni per il trattamento e la patologia delle filariosi.

Filarial nematodes harbour intracellular, Gram-negative bacteria belonging to the genus Wolbachia. These bacteria have been observed in various species of filariae, including the main filariasis agents of humans and animals. It has been suggested that Wolbachia could play an important role in the biology of filarial nematodes and could be implicated in the pathogenesis of filarial diseases. Wolbachia could thus represent a target for the control of filariasis and key to the understanding of these diseases. Indeed, in various species of filariae, tetracycline treatments have been shown both to reduce/eliminate the Wolbachia population and to determine detrimental effects on the nematodes. In addition, proteins of Wolbachia have been shown to determine specific IgG responses in animals infected by filariae and some Wolbachia molecules (e.g. LPS) have been shown to stimulate innate-immunity responses (e.g. production of cytokines such as IL1, IL6, IL10, TNF-alpha and IFN-gamma by macrophages).

Prevalence of Bancroftian filariasis in Burdwan District, west Bengal: a comparative study between colliery and non-colliery areas.

The present study reports results of a survey of bancroftian filariasis in Burdwan district West Bengal. 85,974 persons were examined of which 46,221 were inhabitants of the colliery areas of the district and rest 39,753 were from non-Colliery areas, including urban and rural areas of the district. The clinicopositivity, endemicity, microfilaria(mf)-positivity, mf-clinicopositivity, and mf-density were determined. It was found that all these epidemiological and parasitological parameters were significantly higher in the colliery area as compared to non-colliery area. The high vector density in the colliery area may account for such significantly higher prevalence of filariasis in this area. From a differential count of neutrophils, eosinophils and lymphocytes in the mf-positive cases, it was revealed that neutrophil percentage was significantly (P < 0.01) decreased while eosinophils and Lymphocytes increased. The decrease of neutrophils was correlated ((Correlation co-efficient, r = -0.78) with the increase of lymphocytes, indicating an imbalance in the immune-system of the infected persons. The geographical and geological factors resulting in high vector density are considered responsible for the significantly higher rate of bancroftian filariasis in the Colliery area.