Zoonotic Visceral Leishmaniasis: New Insights on Innate Immune Response by Blood Macrophages and Liver Kupffer Cells to Leishmania infantum Parasites

L. infantum is the aetiological agent of zoonotic visceral leishmaniasis (ZVL), a disease that affects humans and dogs. Leishmania parasites are well adapted to aggressive conditions inside the phagolysosome and can control the immune activation of macrophages (MØs). Although MØs are highly active phagocytic cells with the capacity to destroy pathogens, they additionally comprise the host cells for Leishmania infection, replication, and stable establishment in the mammal host. The present study compares, for the first time, the innate immune response to L. infantum infection of two different macrophage lineages: the blood macrophages and the liver macrophages (Kupffer cells, KC). Our findings showed that L. infantum takes advantage of the natural predisposition of blood-MØs to phagocyte pathogens. However, parasites rapidly subvert the mechanisms of MØs immune activation. On the other hand, KCs, which are primed for immune tolerance, are not extensively activated and can overcome the dormancy induced by the parasite, exhibiting a selection of immune mechanisms, such as extracellular trap formation. Altogether, KCs reveal a different pattern of response in contrast with blood-MØs when confronting L. infantum parasites. In addition, KCs response appears to be more efficient in managing parasite infection, thus contributing to the ability of the liver to naturally restrain Leishmania dissemination.

Inflammatory Nasal Swelling due to Leishmania tropica

Since its discovery in the 19th century, cutaneous leishmaniasis has been a major public health problem, especially with the appearance of more and more unusual cases of cutaneous lesions due to this parasite. Indeed, the present study joins the previous studies and describes a typical case of a nasal lesion due to Leishmania infection. This is a 20-year-old young man, with no particular pathological history, from an epidemic focus who presented with inflammatory nasal swelling similar to a mucocutaneous form. However, the X-ray data showed that no lysis of the bones proper to the nose was detected and no damage to the underlying mucosa was observed. Nevertheless, the parasitological diagnosis confirmed the presence of amastigotes, and the results of the molecular study showed that the electrophoretic profile was comparable to that of L. tropica. After diagnosis and confirmation, treatment with meglumine antimonate at the rate of two ampoules/injection (one ampoule = 5 ml) of antimony salt for one month was administered intramuscularly with favorable outcome. Atypical forms of cutaneous leishmaniasis constitute a rare and unusual entity often leading to diagnostic delay. For this, the clinical examination must take into account both exceptional presentations of Leishmania infection, in particular in subjects living or having stayed in an endemic area, in order to ensure appropriate and early treatment.

The first case of canine visceral leishmaniasis in the southern region of Santa Catarina, an emerging focus of visceral leishmaniasis in Brazil: regional report or reflection of the reality of a country?

Visceral leishmaniasis is among the most neglected diseases in the world and especially affects poor populations of some developing or underdeveloped countries, with human disease being a factor in the occurrence of thousands of deaths every year. Visceral leishmaniasis affect domestic dogs, main reservoirs of the Leishmania parasite in urban environments. Thus, the aim of this study was to analyze the prevalence of canine visceral leishmaniasis in two non-endemic municipalities located in southern Santa Catarina, as well as to identify the main risk factors for infection in dogs. A cross-sectional study was carried out with dogs living in two Zoonoses Control Centers located in the cities of Tubarão and Criciúma. Blood samples were collected and analyzed by real-time PCR for detection of Leishmania infection. Clinical examination and evaluation of the characteristics of the canine population were also carried out. A total of 107 dogs were evaluated, of which 19% were positive for Leishmania, and 85% of the positive cases were asymptomatic. In conclusion, we highlight the detection of cases of canine visceral leishmaniasis in two cities that had not yet reported cases. The high rate of positive asymptomatic animals suggests the maintenance of the parasite in the environment.

Comparative Study of Real-Time PCR (TaqMan Probe and Sybr Green), Serological Techniques (ELISA, IFA and DAT) and Clinical Signs Evaluation, for the Diagnosis of Canine Leishmaniasis in Experimentally Infected Dogs

Canine leishmaniasis (CanL) diagnosis is not fully resolved. Currently, two specific methodologies are in continuous development, the detection of the parasite DNA or RNA in target organs and the detection of specific antibodies against Leishmania sp. For a correct diagnosis, it has been shown that the joint use of this type of test is necessary. In this work, a Sybr Green and a TaqMan Probe based on real time PCRs (qPCR) was performed for the detection of Leishmania sp. in order to correlate the results with clinicopathological and serological evaluations (IFA, ELISA and DAT) to propose an optimal biological sample to be used to detect the parasite in both early and late stages of the infection. A total of four samples were processed: conjunctival swabs, popliteal lymph node aspirates, bone marrow aspirates, and peripheral blood from experimentally infected dogs belonging to a larger study. Our results indicated that a single non-invasive sample (conjunctival swab) and the application of both types of qPCR would be reliable for determining Leishmania infection as well as the disease stage in dogs, thus avoiding bone marrow, lymph node aspirate or blood samples collection.

Metagenomic Analysis Identifying a Rare Leishmania Infection in an Adult With AIDS

Leishmania belongs to a genus of the protozoan parasites that causes leishmaniasis, and includes cutaneous leishmaniasis (CL) and visceral leishmaniasis (VL). In this case, Leishmania amastigotes were found on cytomorphology examination of the bone marrow specimen, followed by 1,076 Leishmania donovani reads using metagenomic next generation sequencing (mNGS). Since being definitely diagnosed with VL/HIV coinfection, the patient was treated with liposomal amphotericin B as the parasite-resistant therapy and was discharged after clinical cure. But nearly a year later, on the mNGS follow-up, L. donovani was detected in the patient’s blood plasma specimen with 941 reads, suggesting that a relapse of leishmaniasis had occurred. These results indicate that leishmaniasis still exists in China and may represent a public health concern. This case could be helpful in the differential diagnosis of leishmaniasis, and for determining disease progression, prevention, and control of vectors and reservoir hosts.

Effects of Leishmania Species on Immune Response against Malaria Parasite in Malaria Leishmania Coinfections

Both malaria and leishmania are most widespread protozoon parasitic diseases, certainly in tropical countries of the world. Malaria leishmania coinfection is common in leishmaniasis endemic areas which is mostly endemic to malaria too. Researchers notice that in cases of malaria leishmania coinfection , leishmania species find the some extent the outcome of malaria infection , but also behavior of malaria parasite species play a significant role to figure the consequences of it. While L. donovani protect from severe malaria complications by suppression of major histocompatibility class Ⅱ , so it diminish the clinical severity of malaria but not malaria parasite density due to dysfunction of major histocompatibility class I, which controlled by suppressed one, In another side L. mexicana tends to sequester in macrophages and lead to severe clinical outcomes when it coexisted with malaria parasite at same host. Experimental studies required to know more information about coinfection of different malaria and leishmania species to establish clinical research. Leishmania infection excluded when studies aim to assess the immune response to only malaria parasite, experimental studies required involving different species of malaria and leishmania.

Detection of Leishmania donovani in Wild-Caught Phlebotomine Sand Flies in Endemic Focus of Leishmaniasis in Himachal Pradesh, India

An endemic focus of cutaneous leishmaniasis (CL) is located in the Sutlej River Valley in Himachal Pradesh (India) in the north-western Himalaya where CL co-exists with visceral leishmaniasis (VL). In areas of the Indian subcontinent such as Rajasthan, cutaneous leishmaniasis is transmitted by Phlebotomus papatasi (Scopoli) and Phlebotomus salehi. In Himachal Pradesh, Phlebotomus longiductus (Parrot) is suspected to be the vector for CL. In the current study, sand flies were collected and tested for Leishmania infection and to confirm the vector species causing CL. Sand flies were collected during April and September of 2017–2019 from CL endemic villages of Rampur (Shimla), Nirmand (Kullu) and Nichar (Kinnaur) districts of Himachal Pradesh. The sand flies were identified as Phlebotomus (adlerius) longiductus (Parrot) and Phlebotomus (larrousisus) major (Annandale). The density of P. longiductus was found highest. The elevation of villages ranged from 947 m to 2,130 m and were far from the presence of subsoil water. Field collected sand flies tested positive (7.69%) for Leishmania donovani by PCR-RFLP. The L. donovani sequences detected from P. longiductus were 97% similar to L. donovani sequences reported from the cases of CL in Himachal Pradesh.The Leishmania positive sand flies were morphologically identified as Phlebotomus adlerious longiductus providing one step further evidence towards the vector status of CL in Himachal Pradesh. The findings of the study are of epidemiological significance for strategic planning of vector control for leishmaniasis in India.

Leishmania Infection during Chemotherapy in a Dog Diagnosed with Multicentric Large B-Cell Lymphoma—A Diagnostic Challenge

Dogs with lymphoma are at risk of developing clinical complications due to immunosuppression and side effects of chemotherapy. Clinical reports of concurrent lymphoma and leishmaniasis are rare and confined to single cases of comorbidity at presentation. Herein, we describe a case of lymphoma during maintenance chemotherapy in which bone marrow cytology showed myelodysplasia associated with leishmaniasis. The dog was a seven-year-old intact female Parson Russel Terrier with a two-week history of generalized lymphadenopathy. Diagnosis of multicentric high-grade B-cell lymphoma stage Va was carried out with cytological and cytofluorimetric assays of external lymph nodes, abdominal ultrasound, chest radiology, and lymphoid blasts blood smear examination. The dog lived and had traveled in endemic areas of Leishmania with uninterrupted prevention against sand fly bites by an insecticide-impregnated collar and presented seronegativity to Leishmania at presentation. Chemotherapy for lymphoma was successful and the patient achieved complete remission. Approximately eight months after the diagnosis, a persistent pancytopenia was assessed. Unexpectedly, Leishmania amastigotes were identified in the bone marrow. Combined treatment rounds were administered with antileishmanial and antineoplastic drugs for approximately eight months. Eventually, lymphoma relapsed and became unresponsive to chemotherapy, and the dog was euthanatized. Canine lymphoma overlapping with subsequent Leishmania infection as a complication is rare and lacks specific clinical manifestations. A delayed diagnosis of leishmaniasis may occur. We suggest considering leishmaniasis as part of the differential diagnosis of persistent pancytopenia in dogs with lymphoma, particularly in dogs who reside or travel to endemic areas, when treatment fails or abnormal laboratory findings are present.

Leukocyte Behavior in Mesenteric Microcirculation upon Exper-imental By Leishmania Spp. in BALB/c Mice

Background: We aimed to determine the cellular recruitment (leukocyte rolling and adhesion) by which the Leishmania (Viannia) braziliensis, L. (Leishmania) amazonensis, and L. (Leishmania) major species in the mesenteric microcirculation of BALB/c mice. Methods: Five experimental groups were considered: group 1 (L. braziliensis); group 2 (L. amazonensis); group 3 (L. major); group 4 (control group with PBS); group 5 (negative control group), analyzed 3, 6, 12, and 24 h after parasite inoculation. Results: Infections by the different Leishmania species caused an increase in the number of rolling leukocytes: L. braziliensis a peak at 6 h; L. amazonensis and L. major a peak at 3 h. The Leishmania infections induced leukocyte adhesion: L. major and L. amazonensis showed an increase after 3 and 6 h, respectively. Conclusion: The kinetics of cellular recruitment in Leishmania infections, leading to infection susceptibility or resistance, indicates that distinct mechanisms regulate the initial response to Leishmania infection and determine its course.

Mast Cell Response to Leishmania mexicana and Sand Fly Salivary Proteins is Modulated by Androgens

Mast cells (MCs) play a crucial role during infections with Leishmania, that is transmitted through the bite of an infected sand fly that injects saliva together with the parasite. Sand fly saliva is a complex fluid that modulates the host immune response. In addition, hormonal factors modulate the host immune response, impacting the susceptibility to infections. Thus, to assess the impact of androgens and salivary proteins of sand fly vectors on the mast cell (MC) response to Leishmania infections, we infected orchiectomized male mice with the parasite in the presence or absence of sand fly salivary proteins and analyzed the inflammatory response of MCs. Our results showed a differential MC response to the parasite and to vector salivary proteins in mice deprived of gonadal hormones, as compared to sham-operated mice. Orchidectomy induced a different pattern of activation in MC of animals infected with Leishmania and vector-salivary proteins. Our results show that during Leishmania infection, androgens modulate the innate immunity response against the parasite and salivary proteins of the sand fly vector.