What is best marker to analyze macrophage, neutrophils and plasma cells in parasite (Trematode) induced granuloma in the human eye by immunohistochemistry methods?
Best marker for human granulocytes is CD66b (CEACAM8). It has a long standing history and is generally used in clinic. As GPI-linked molecule it is used for diagnosis of Paroxysmal nocturnal hemoglobinuria (if you have it CD66b is negative). CD66b is solely expressed in human PMN and not in any other leukocyte subpopulation and not in epithelia and endothelia. Furthermore it can be sued as an activation marker for human PMN because it is up-regulated within minutes after stimulation with fMLP, PMA etc. Due to its presence in neutrophilic vesicles. CD66b is present in neutrohilic, acurophilic and basophilic human granulocytes. In cas of working with rat and mouse system you may consider to use CD66a (CEACAM1) although it is present in lymphocytes but there with extremely low expression level.
Hi Lalan,
The best marker for human neutrophils for immunohistochemistry is anti-human neutrophil elastase. Neutrophil elastase is only expressed in neutrophils (found in the azurophilic granules) and not in other cells. Another excellent marker is Ly6G (using clone 1A8), which is found in the plasma membrane of neutrophils.
I'm not sure what is best for macrophages but would suggest CD14, CD68, and/or EMR antibody. A quick search on Google shows that CD28 is a good marker for human plasma cells.
Hope this helps!
Paige
I agree with Paige about Neutrophil elastase for human PMNs and CD68 for human macrophages. However, I am not sure that human PMNs express Ly6G. As I know Ly6G is nice marker only for mouse PMNs.
Neutrophil elastase is good marker for for human PMNs, cd14 and cd68 for macrophages and cd28 for plasma cells. lalitha kabilan
CD14 is on both macrophages and neutrophils (though at different levels). MPO for neuts?
Thanks for the clarification, Rinat - you are correct, Ly6G is only for mouse PMNs. I get my species mixed up once in a while!
MPO is a good marker for purified neutrophils but is also present in monocytes/macrophages, so is not so ideal as a tissue marker.
I agree with Paige about markers for Neutrophil(elastase) and human macrophages(CD68). However, CD28 is a receptor expressed on T cell that provide co-stimulating signal for activation. Therefore, I suggest that CD138 is a good choice for human plasma cell.
syndecan-1 for plasma cells, at least in in mice, CD138-but very hi IgG would also be suitable in some cases, and they may still have some CD19
Best marker for human granulocytes is CD66b (CEACAM8). It has a long standing history and is generally used in clinic. As GPI-linked molecule it is used for diagnosis of Paroxysmal nocturnal hemoglobinuria (if you have it CD66b is negative). CD66b is solely expressed in human PMN and not in any other leukocyte subpopulation and not in epithelia and endothelia. Furthermore it can be sued as an activation marker for human PMN because it is up-regulated within minutes after stimulation with fMLP, PMA etc. Due to its presence in neutrophilic vesicles. CD66b is present in neutrohilic, acurophilic and basophilic human granulocytes. In cas of working with rat and mouse system you may consider to use CD66a (CEACAM1) although it is present in lymphocytes but there with extremely low expression level.
CD66b is also expressed in eosinophils so is not unique to neutrophils. There may be a few eosinophils in the human eye during parasite-induced infections. Miltenyi has a nice description of the expression and function of CD66b: http://www.miltenyibiotec.com/en/products-and-services/macs-flow-cytometry/reagents/antibodies-and-dyes/cd66b-antibodies-human.aspx
For human plasma cells we use CD45 (-) CD 38 (+), CD 138 (+) and CD 56 (+) -for malignant cells!
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