Cell lines infected with granulocytic ehrlichia, vaccines, diagnostics and methods

A method of continually growing human granulocytic Ehrlichia, E. equi, or E. phagocytophila comprising

• a) infecting a cell line, wherein the phenotype of the cell line is selected from the group consisting of a promelocytic leukemia cell line, an acute myelogenous leukemia cell line, a histiocytic lymphoma cell line, an acute monocytic leukemia cell line, and an embryonic lung cell line, with human granulocytic Ehrlichia, E. equi, or E. phagocylophila by co-culturing the cell line with blood cells infected with human granulocytic Ehrlichia, E. equi, or E. phagocytophila within a suitable culture medium;
• b) culturing the infected cell line with fresh uninfected cultured cells;
• c) splitting the infected cell line in a split ratio of about 1:2 or 1:3 of infected cells to additional said fresh uninfected cultured cells in order to maintain the culture;
• and
• d) identifying a cytopathic effect on the infected cell line, wherein the means for identifying the cytopathic effect comprise
• i) using an immunofluorescence assay, Western blot analysis, EIA or RIA to detect the presence of human granulocytic Ehrlichia, E. equi or E. phagocytophila using detector antibodies specific for said human granulocytic Ehrlichia, E. equi or E. phagocytophila;
• ii) using PCR or RFLP to confirm the presence or absence of human granulocytic Ehrlichia, E. equi or E. phagocytophila in the cell lines with a primer specific or a 16S rDNA gene specific for said human granulocytic Ehrlichia, E. equi or E. phagocytophila;
• or
• iii) observing degenerative cell morphology by light microscopy.