Adoptive Transfer and CD4 Depletion in Leishmaniasis

Spleen cells from C57BL/6 mice were collected, erythrocytes lysed with ACK lysing buffer (Quality Biological, Cat #118-156-101), and CD8⁺ T cells were purified using a magnetic bead separation kit (Miltenyi Biotec, Cat #130-104-075). Rag1^−/− mice were reconstituted with CD8⁺ T cells by intravenous route (3 × 10⁶ cells/mouse) and subsequently were infected with L. braziliensis. Mice received 250 mg of antibody anti-CD4, clone GK1.5 (BioXCell, Cat #BE0003-1, RRID:AB_1107636) by intraperitoneal injections twice a week in the first two weeks post-infection[8 (link)].

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Sacramento L.A., Amorim C.F., Lombana C.G., Beiting D., Novais F., Carvalho L.P., Carvalho E.M, & Scott P. (2023). CCR5 promotes the migration of CD8+ T cells to the leishmanial lesions. bioRxiv.

Publication Preprint 2023

ACK lysing buffer magnetic bead separation kit

Anti antibody Biological Buffer C57bl mice Cat 1 Cd8 t cells Cells Clone Erythrocytes Infection Intraperitoneal injections Mice Rag1 Spleen

Corresponding Organization : University of Pennsylvania

Other organizations : The Ohio State University, Complexo Hospitalar Universitário Professor Edgard Santos, Universidade Federal da Bahia

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Variable analysis

independent variables

Reconstitution of Rag1-/- mice with CD8+ T cells
Infection of reconstituted Rag1-/- mice with L. braziliensis
Administration of anti-CD4 antibody (clone GK1.5)

dependent variables

Not explicitly mentioned

control variables

Mouse strain (C57BL/6)
Rag1-/- mice
Intravenous route of CD8+ T cell reconstitution
Dose of CD8+ T cells (3 x 10^6 cells/mouse)
Timing and route of anti-CD4 antibody administration (twice a week for the first two weeks post-infection, intraperitoneal)

controls

Positive control: None mentioned
Negative control: None mentioned

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