0000000000040090
AUTHOR
Michael R. Fischer
Humanized mice in cutaneous leishmaniasis—Suitability analysis of human PBMC transfer into immunodeficient mice
Humanized mice represent a suitable preclinical test system for example therapeutic interventions in various disease settings, including infections. Here, we intended to establish such system for cutaneous leishmaniasis by infecting T, B and NK cell-deficient mice adoptively transferred with human peripheral blood mononuclear cells (PBMC). L major infection led to the establishment of parasite lesions harbouring viable parasites and human T cells, but parasite elimination was not seen due to a species-specific activity of T cell-derived human IFNγ. In addition, up to 50% of infected mice succumbed to severe graft-versus-host disease. In summary, even though long-term disease outcome assessm…
Blue light irradiation suppresses dendritic cells activationin vitro
Blue light is a UV-free irradiation suitable for treating chronic skin inflammation, for example, atopic dermatitis, psoriasis, and hand- and foot eczema. However, a better understanding of the mode of action is still missing. For this reason, we investigated whether dendritic cells (DC) are directly affected by blue light irradiation in vitro. Here, we report that irradiation neither induced apoptosis nor maturation of monocyte-derived and myeloid DC. However, subsequent DC maturation upon LPS/IFNγ stimulation was impaired in a dose-dependent manner as assessed by maturation markers and cytokine release. Moreover, the potential of this DC to induce cytokine secretion from allogeneic CD4 T …
Immune Modulating Effects of NKT Cells in a Physiologically Low Dose Leishmania major Infection Model after αGalCer Analog PBS57 Stimulation
Leishmaniasis is a parasitic infection affecting ∼12 million people worldwide, mostly in developing countries. Treatment options are limited and no effective vaccines exist to date. Natural Killer T (NKT) cells are a conserved innate-like lymphocyte population with immunomodulating effects in various settings. A number of reports state a role of NKT cells in different models of Leishmania infection. Here, we investigated the effect of NKT cells in a physiologically relevant, intradermal low dose infection model. After inoculation of 103 infectious-stage L. major, comparable numbers of skin-immigrating NKT cells in both susceptible BALB/c mice and resistant C57BL/6 mice were noted. Compared …
Myeloid cells do not contribute to gender-dependent differences in disease outcome in murine cutaneous leishmaniasis.
Gender-associated differences in the outcome of infections are well known. Apart from behavior-released differences in their incidence, immunological factors also contribute to disease outcome. The underlying mechanisms are often unknown. Here, we show that in murine experimental leishmaniasis, female mice develop larger skin lesions that harbor significantly more parasites, exhibit increased parasite dissemination to visceral organs associated with a shift towards T helper (Th) 2 immunity with increased levels of IL-4. Antigen presenting cells (APC) responsible for T cell priming, such as macrophages or dendritic cells, were not involved in the process. Additionally, in adoptive transfer e…
Animal Model for Cutaneous Leishmaniasis
Using cutaneous leishmaniasis of mice, the existence of so-called T helper (Th) cells type 1 and type 2 had been identified more than 20 years ago. Nowadays, it is well accepted that additional T cell populations as well as B cell-mediated immunity is required for immunity against Leishmania major. Finally, using inbred mouse strains, the relevance of genetical factors that influence anti-pathogen immunity as well as elements of the skin-immune system have been identified. This protocol describes a model for murine experimental leishmaniasis that tries to mimic natural parasite transmission by several means: (1) utilization of only infectious-stage parasites that are found in sand fly saliv…
The Dissociation Constant of Pyruvic Acid: Determination by Spectrophotometric Measurements
Aqueous solutions of sodium pyruvate/hydrochloric acid mixtures were studied with regard to changes in the intensity of optical absorption at 317 nm wavelength, caused by varying pH and temperature. The data were evaluated to determine the pyruvic acid apparent dissociation constant at infinite dilution. Its temperature dependence followed the relation ln Kod = −(0.55 ± 0.47) − (1538 ± 135)/T or, if previous results of Pedersen at two temperatures are included, In Kod = −(0.97 ± 0.35) − (1418 ± 102)T. The effective absorption coefficient for pyruvic acid and its anion differ because the former species is largely hydrated, whereas the latter is not. The known hydration constant and its tempe…
Model for Generation of Large Numbers of Primary, Inflammatory Skin-Derived Neutrophils, and Macrophages
Tissue macrophages and inflammatory neutrophils represent important cells of the innate immune system responsible for various important tasks, i.e., elimination of pathogens and/or granuloma formation. Isolation of large numbers of primary phagocytes is vital for research with these cells. Within this protocol, we present a strategy for isolation of large numbers of inflammatory neutrophils and macrophages from murine skin that allows for follow-up in vivo or in vitro studies.