US 9,175,334 B2Granted

Identifying viral cell tropism

AssigneesCentre National De La Recherche Scientifique, Assistance Publique-Hopitaux de Paris, Universite Montpellier 2 Sciences Et Techniques, Universite Paris Diderot—Paris 7

InventorsCharles-Henri Lecellier, Valerie Courgnaud, Manuella Bouttier, Diane Descamps, Gilles Collin

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Abstract

The invention relates to an in vitro method for identifying microRNAs or the target mRNAs thereof, the expression of which during the infection of cells by a virus using a cell receptor and at least one cell co-receptor for entering the cell, is specifically modified on the basis of the cell co-receptor used by the virus for its entering the cells, comprising: i) determining the microRNA expression levels in a test cell expressing a receptor, a first co-receptor and at least one other co-receptor, after infection by a first virus using the first co-receptor and by at least one other virus using another co-receptor, respectively; ii) identifying the microRNAs, the expression level of which is modulated during the infection by each of the viruses in relation to the expression level in the uninfected cells; iii) comparing the thus-identified microRNAs; iv) selecting the microRNAs, the modification of the expression level of which is specific to the use of a co-receptor; v) optionally identifying the target mRNAs of the thus-selected microRNAs.

Claims6

Dependent claims

An in vitro method for determining if a cell co-receptor is used by a virus using a cell receptor and at least one cell co-receptor for entering a cell, in a patient infected with the virus, comprising:

i) putting a sample of the patient which contains the virus in contact with a test cell expressing a cell receptor of the virus and at least one cell co-receptor of the virus, wherein the use of said cell co-receptor by the virus specifically modulates the expression of at least one miRNA;
ii) determining the expression level of said at least one miRNA and/or of at least one target mRNA of said at least one miRNA in the test cell;
iii) comparing the expression level with a predetermined value, said predetermined value being:
the expression level of said at least one miRNA and/or of said at least one target mRNA in a cell expressing said at least one co-receptor and infected with a reference virus using said at least one co-receptor for entering the cell, or
the expression level of said at least one miRNA and/or of said at least one target mRNA in an uninfected cell expressing said at least one co-receptor;
and
iv) inferring therefrom whether the virus uses or not said at least one cell co-receptor expressed by said test cell,
wherein the virus is the HIV virus, and wherein the test cell expresses the cell receptor CD4 and a cell co-receptor selected from the group consisting of CXCR4, CCR5, CCR3, CCR2, CCR1, CCR4, CCR8, CCR9, CXCR2, STRL33, V28, gpr1, gpr15 and ChemR23, and wherein said at least one miRNA is selected from the group consisting of hsa-miR574-5p, hsa-miR-663, hsa-miR-149*, hsa-miR-575, hsa-miR-638, hsa-miR-181b, hsa-let-7g, hsa-miR-30a, hsa-miR-148a and hsa-miR-9*.

5 more elements

Dependencies are inferred from the claim text.