Inherent biomechanical traits enable infective filariae to disseminate through collecting lymphatic vessels.

Filariases are diseases caused by arthropod-borne filaria nematodes. The related pathologies depend on the location of the infective larvae when their migration, the asymptomatic and least studied phase of the disease, comes to an end. To determine factors assisting in filariae dissemination, we image Litomosoides sigmodontis infective larvae during their escape from the skin. Burrowing through the dermis filariae exclusively enter pre-collecting lymphatics by mechanical disruption of their wall. Once inside collectors, their rapid and unidirectional movement towards the lymph node is supported by the morphology of lymphatic valves. In a microfluidic maze mimicking lymphatic vessels, filariae follow the direction of the flow, the first biomechanical factor capable of helminth guidance within the host. Finally, non-infective nematodes that rely on universal morpho-physiological cues alone also migrate through the dermis, and break in lymphatics, indicating that the ability to spread by the lymphatic route is an ancestral trait rather than acquired parasitic adaptation.

A lymphatic dwelling filarioid nematode, Rumenfilaria andersoni (Filarioidea; Splendidofilariinae), is an emerging parasite in Finnish cervids.

BACKGROUND: Recent studies revealed expansion of filarioid nematodes into northern Finland. In addition to Setaria tundra, an abundant filarioid, Rumenfilaria andersoni, was found inhabiting the lymphatic vessels of reindeer. Our study explores the dynamics of the rapid geographic expansion of R. andersoni, defining prevalence and density of microfilariae among 4 new cervid host species in Finland while developing a context for host-parasite ecology in Fennoscandia and more broadly in the Arctic and boreal regions. METHODS: Blood samples were evaluated for presence of microfilariae from 1576 semi-domesticated reindeer, 8 captive reindeer, and free-ranging cervids including 105 wild forest reindeer, 862 moose, 114 white tailed deer and 73 roe deer in 2003-2006 (-2010). Additionally, the prepatent period and the efficacy of ivermectin treatment were investigated. RESULTS: Rumenfilaria andersoni was found to be a common and abundant parasite in reindeer (0-90%) and wild forest reindeer (41-100%). Also moose (0-12%), white-tailed deer (15-22%) and roe deer (3%) were revealed as definitive hosts. Ivermectin was not effective against adult parasites. The prepatent period was estimated to be about five months. CONCLUSIONS: Rumenfilaria andersoni was identified in 3 endemic cervid species and the introduced white-tailed deer, all constituting previously unrecognized host species in the Palearctic. Among moose, the prevalence and intensity were substantially lower than levels observed among subspecies of reindeer. White-tailed deer had a relatively high prevalence and density of R. andersoni microfilariae (rmf), whereas our limited data for roe deer indicated that the nematode may not have been abundant. Density and prevalence of rmf in moose and white tailed deer suggests the nematode may be adapted to these species, and that these cervids may be among the primary hosts of R. andersoni and reservoirs for transmission in Finland. Our current data suggest that R. andersoni became established in Finland recently, coincidental with introduction of white-tailed deer from North America in 1935; subsequent invasion and emergence in the past 70-80 years appears driven by climate-related factors. An alternative hypothesis for a temporally deeper occurrence for R. andersoni in Fennoscandia, representing post-Pleistocene range expansion with moose tracking deglaciation, is not firmly supported.

Lymphatic dwelling filarioid nematodes in reindeer Rangifer tarandus tarandus, (cervidae) in Finland, identified as Rumenfilaria andersoni Lankester & Snider, 1982 (nematoda: Onchocercidae: Splendidofilariinae).

A filarioid nematode inhabiting the lymphatic vessels of the subserosal rumen and mesenteries associated with a high prevalence of its microfilariae in peripheral blood was observed in Finnish reindeer (Rangifer tarandus tarandus) in 2004 and 2006. Adult specimens were collected by dissecting lymphatic vessels from slaughtered animals, where some of the nematodes were seen through the wall of the dilated vessels as thin white winding threads obscuring the vessel. The morphology of adult worms and microfilaria is described based on light and scanning electron microscopy. These filariae belong to the subfamily Splendidofilariinae of the Onchocercidae and resemble Rumenfilaria andersoni, recovered from different host and localization, the ruminal veins of Alces alces in Canada. Comparison of paratypes of this species revealed only minor differences which were not sufficient to separate the filarioid parasitic in R. tarandus in Finland and we identify the nematode as R. andersoni. However, the findings suggest two different parasite populations. The finalizing of this taxonomic question in the future requires an integrated approach, in which the DNA-based and morphological identifications are consistent.

The association of adult Onchocerca volvulus with lymphatic vessels.

Immunocytochemical examination of onchocercal nodule tissues containing adult Onchocerca volvulus using immuno-markers for blood and lymphatic vessels (vWF, D2-40, podoplanin, Prox-1, and Lyve1) shows a distinct pattern of distribution of these vessels within nodules. Blood vessels were commonly seen associated with organized lymphoid cellular aggregates in the both the outer and inner areas of the nodules. In contrast, the majority of the lymphatic vessel positivity was seen in the central zone in close apposition to the adult parasites, and the remainder usually associated with microfilariae in the outer areas of the nodule. These findings suggest an intimate relationship between adult O. volvulus and lymphatic vessels, including the likely proliferation of lymphatic endothelial cells (lymphangectasia) akin to that seen with other filariae. These findings indicate that adult O. volvulus may migrate via the lymphatic system, and that clinical manifestations of this disease that involve tissue edema may be the result of the location of these worms in the lymphatic system.

Lymphatic vascularisation and involvement of Lyve-1+ macrophages in the human onchocerca nodule.

Onchocerciasis, caused by the filarial nematode Onchocerca volvulus, is a parasitic disease leading to debilitating skin disease and blindness, with major economic and social consequences. The pathology of onchocerciasis is principally considered to be a consequence of long-standing host inflammatory responses. In onchocerciasis a subcutaneous nodule is formed around the female worms, the core of which is a dense infiltrate of inflammatory cells in which microfilariae are released. It has been established that the formation of nodules is associated with angiogenesis. In this study, we show using specific markers of endothelium (CD31) and lymphatic endothelial cells (Lyve-1, Podoplanin) that not only angiogenesis but also lymphangiogenesis occurs within the nodule. 7% of the microfilariae could be found within the lymphatics, but none within blood vessels in these nodules, suggesting a possible route of migration for the larvae. The neovascularisation was associated with a particular pattern of angio/lymphangiogenic factors in nodules of onchocerciasis patients, characterized by the expression of CXCL12, CXCR4, VEGF-C, Angiopoietin-1 and Angiopoietin-2. Interestingly, a proportion of macrophages were found to be positive for Lyve-1 and some were integrated into the endothelium of the lymphatic vessels, revealing their plasticity in the nodular micro-environment. These results indicate that lymphatic as well as blood vascularization is induced around O. volvulus worms, either by the parasite itself, e.g. by the release of angiogenic and lymphangiogenic factors, or by consecutive host immune responses.

Quantitative imaging of Plasmodium transmission from mosquito to mammal.

Plasmodium, the parasite that causes malaria, is transmitted by a mosquito into the dermis and must reach the liver before infecting erythrocytes and causing disease. We present here a quantitative, real-time analysis of the fate of parasites transmitted in a rodent system. We show that only a proportion of the parasites enter blood capillaries, whereas others are drained by lymphatics. Lymph sporozoites stop at the proximal lymph node, where most are degraded inside dendritic leucocytes, but some can partially differentiate into exoerythrocytic stages. This previously unrecognized step of the parasite life cycle could influence the immune response of the host, and may have implications for vaccination strategies against the preerythrocytic stages of the parasite.

Profiling the cellular immune response to multiple Brugia pahangi infections in a susceptible host.

Human lymphatic filariasis is caused primarily by Brugia malayi and Wuchereria bancroffi. Unraveling this disease is complex, as people living in endemic areas exhibit a vast array of clinical states and immune responses. The Mongolian gerbil (Meriones unguiculatus)-B. pahangi model of human lymphatic filariasis has provided much information on immune parameters associated with filarial infection. Prior investigations in our laboratory have shown that gerbils closely mimic a subset of patients classified as microfilaremic but asymptomatic, a group that comprises the majority of people living in endemic areas. Worm recovery data suggest that gerbils carrying current B. pahangi infections do not show any resistance to subsequent subcutaneous B. pahangi infections. The aim of the present studies was to investigate the T cell cytokine response in gerbils receiving multiple infections of B. pahangi as a means of mimicking the conditions experienced by people in endemic areas. The T cell cytokine profile generated by multiply infected gerbils was not different from that previously generated by gerbils infected only once with B. pahangi. Gerbils infected multiple times with B. pahangi showed a transient increase in IL-5, which corresponded to the increased eosinophil levels previously reported from multiply infected gerbils. Chronically infected gerbils showed elevated IL-4 mRNA levels, as has been reported from gerbils infected only once with B. pahangi. Chronic infections were also associated with a state of immune hyporesponsiveness, as determined by the characterization of lymphatic thrombi and lymphoproliferation of spleen and renal lymph node cells to worm antigen.

Reliable and frequent detection of adult Wuchereria bancrofti in Ghanaian women by ultrasonography.

Detection of adult Wuchereria bancrofti by ultrasonography of the scrotal region in men is a suitable diagnostic tool for lymphatic filariasis, whereas there are only a few case reports of adult filariae observed by ultrasonography in women. We examined 35 microfilaraemic women ultrasonographically in sites of the body suspected as locations for worm nests. In 15 women the 'filaria dance sign' (FDS) was detected in various locations, some being novel, such as adult worms within lymphatic vessels between muscular fibres of the thighs. The surprisingly high number of worm nests detectable in microfilaraemic women recommends ultrasonography for diagnosis and treatment efficacy monitoring of female patients infected with W. bancrofti.

An inguinal mass with local vascular lesions induced by a lymphatic filaria.

A 47-year-old Indian male presented with an inguinal mass clinically suspicious as a tumor. Histological examination of the excised mass demonstrated tissue reaction to degenerating intravascular adult filarial worms. The worms have been identified as a lymphatic filariae, most probably Wuchereria bancrofti. The case report underscores the need to maintain suspicion of genitourinary filarial lesions in non-endemic areas and describes atypical vascular lesions induced by lymphatic filariae.