IOVS Human Reproduction
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(Investigative Ophthalmology and Visual Science. 2008;49:1010-1015.)
© 2008 by The Association for Research in Vision and Ophthalmology, Inc.
DOI:  10.1167/iovs.07-1130
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Milk Components Inhibit Acanthamoeba-Induced Cytopathic Effect

Chandrassegar Saravanan,1,2,3 Zhiyi Cao,2,3 Janardan Kumar,2,4 Jiazhou Qiu,5 Andrew G. Plaut,5 David S. Newburg,6 and Noorjahan Panjwani1,2

1From the Department of Anatomy and Cell Biology, Sackler School of Graduate Biomedical Sciences, 2Department of Ophthalmology, Center for Vision Research and The New England Eye Center, Tufts University School of Medicine, Boston, Massachusetts; 5Tufts-New England Medical Center, Boston, Massachusetts; and 6Program in Glycobiology, Pediatric Gastroenterology and Nutrition, Massachusetts General Hospital, Charlestown, Massachusetts.

PURPOSE. Acanthamoebae provoke a vision-threatening corneal infection known as Acanthamoeba keratitis (AK). It is thought that Acanthamoeba-specific IgA antibodies present in mucosal secretions such as human tears, milk, and saliva provide protection against infection by inhibiting the adhesion of parasites to host cells. The goal of the present study was to determine whether human mucosal secretions have the potential to provide protection against the Acanthamoeba-induced cytopathic effect (CPE) by an additional mechanism that is independent of IgA.

METHODS. Breast milk was used as a model of human mucosal secretions. In vitro CPE assays were used to examine the CPE inhibitory effect of IgA-depleted milk and various milk fractions obtained by gel filtration. The activity of amebic proteinases was examined by zymography.

RESULTS. IgA-depleted milk inhibited the Acanthamoeba-induced CPE in a concentration-dependent manner. Milk proteins were separated into four major fractions (F1-F4) by gel filtration. Of these four fractions, CPE inhibitory activity was detected largely in fraction F3. In contrast, fractions F1, F2, and F4 lacked CPE inhibitory activity. Moreover, fraction F3, but not F1, F2, or F4, inhibited amebic proteinases.

CONCLUSIONS. These data, in conjunction with published findings showing that amebic proteinases are responsible for the induction of Acanthamoeba CPE, led us to propose that human mucosal secretions have the potential to provide protection against Acanthamoeba-induced CPE by an additional mechanism that is independent of IgA and that involves the inhibition of cytotoxic proteinases of amebae.






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