Geographical Distribution of MDR1 Expression in Leishmania Isolates, from Greece and Cyprus, Measured by the Rhodamine-123 Efflux Potential of the Isolates, Using Flow Cytometry

Abstract

Abstract. Leishmaniasis, a neglected vector-borne disease caused by the protozoan parasite Leishmania, is encountered in 98 countries causing serious concerns to public health. The most alarming is the development of parasite drug resistance, a phenomenon increasingly encountered in the field rendering chemotherapy ineffective. Although resistance to drugs is a complex phenomenon, the rate of efflux of the fluorescent dye Rhodamine-123 from the parasite body, using flow cytometry, is an indication of the isolate's ability to efflux the drug, thus avoiding death. The rate of efflux measured 275 Leishmania strains, isolated from patients and dogs from Greece and Cyprus, was measured and mapped to study the geographical distribution of the multidrug resistance (MDR) gene expression as an indication of the drug resistance of the parasite. The map showed that out of the seven prefectures, where dogs presented high efflux rates, five also had patients with high efflux rates. In one, out of the 59 prefectures studied, the highest number of isolates with efflux slope α > 1, in both human and dog isolates, was found; a fact which may suggest that spread of drug resistance is taking place. The virulence of the Leishmania strains, assessed after infecting human macrophages of the THP-1 cell line, fluctuated from 1% to 59.3% with only 2.5% of the isolates showing infectivity > 50%. The most virulent strains were isolated from Attica and Crete.

The Current Status of Protozoan Parasitic Diseases in Cyprus: A Narrative Literature Review

“…The data obtained from the questionnaire was compared with data from two recent epidemiological studies on leishmaniasis carried out in Greece and Cyprus at the same time using statistical methods and spatial statistics. Tsirigotakis et al (2016) [10] analyzed the multidrug resistance gene (MDR1) expression in Leishmania species from Greece and Cyprus and represented it geographically to assess the distribution of drug resistance in the region. They tested the rate of efflux of the fluorescent dye Rhodamine-123 (Rhod-123) by flow cytometry in 211 Leishmania strains isolated from patients and dogs in Greece and 64 strains isolated from patients and dogs in Cyprus.…”

High resolution melting analysis and detection of Leishmania resistance: the role of multi drug resistance 1 gene

“…For instance, Alizadeh and colleagues by using RFLP-PCR method illustrated that about 11% of the collected samples of cutaneous leishmaniasis from some endemic areas of Iran (Yazd, Mashhad, and Kashan provinces) had mutations in MDR1 gene [ 36 ]. Tsirigotakis et al also evaluated drug resistance in leishmania isolates using flow cytometry [ 37 ]. Since Rhodamine-123 is an established substrate for P-gp 170 and its efflux rate is largely dependent on the number of efflux pumps, they introduced the efflux rate of this dye as an indicator of drug resistance.…”