In HSV-1 plaque assay, how much of pooled human immunoglobulin is enough to block secondary infection through the medium? I've found two HSV-1 plaque assay protocols with IgG, one recommending an IgG concentration of 32 ug/ml, the other only 7.5 ug/ml. Does the serum concentration in the upkeep medium affect the sufficient IgG concentration?
We have done this many years ago, but the neutralization effect may be never complete. In addition, the virus spreads from cell to cell, thus the effect may be different depending of the density of cells. Thus, the issue seems more complicated as it looks at first glance.
Thank you very much for your answer! So did you end up using an overlay method such as agarose or methylcellulose instead of IgG?
Not at all. Adding virus specific IgG in our hands seemed reasonable only in special experiments, such as to mimic the estaglishment of latency in cells infected at low multiplicity and then keeping them for sevedral passages in the presence of antibody. The question was what happens if the antibody was later on withdrawn. Another experiments were made with the rabbit or murine trigeminal ganglia in which latency had been established in vivo. The question was whether reactivation could be prevented. in antibody presence. Even though were was a considerable difference as compared to ganglion fragemnts kept in the absence of IgG, the reactivation in its presence was never fully abolished.
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