TABLE 1.

Experimental protocol for T-cell depletion experiments

Micea	Primary inoculationb	T-cell depletionc	Collect tissued	Vaginal washe	Illness scoresf
Nonimmune	No	No	5 Yes	No	No
			5 No	Yes	Yes
Immune	Yes	No	10 Yes	No	No
			10 No	Yes	Yes
Immune depleted	Yes	Yes	10 Yes	No	No
			10 No	Yes	Yes

^aFifty age-matched females were divided into three groups: 10 nonimmune, 20 immune, and 20 immune depleted. All were treated with 10 μg of estradiol benzoate in peanut oil subcutaneously followed 1 day later by 2.0 mg of Depo-Provera (Upjohn Co., Kalamazoo, Mich.) in PBS subcutaneously 5 days prior to primary inoculation. (The hormone treatment induces susceptibility to vaginal HSV-2 infection [16].) The hormone treatment was repeated 6 days prior to challenge. All mice were subjected to a vaginal wash for IgG anti-HSV-2 either 1 day prior to challenge or on the day of challenge and were challenged at 6 weeks after primary inoculation, using IVAG inoculation of 20 μl of TK⁺HSV-2 at 10⁷ PFU/ml into anesthetized mice. 

^bImmunization by IVAG inoculation of 20 μl of ΔTK⁻HSV-2 at 1.7 × 10⁶ PFU/ml into mice anesthetized with tribromoethanol. 

^cSeven days prior to challenge, using intraperitoneal injection of 0.35 ml of anti-CD8 ascites fluid, 0.9 ml of anti-Thy-1.2 ascites fluid, and 0.3 ml of anti-CD4 ascites fluid followed by another 0.3 ml at 3 days prior to challenge. 

^dVagina and blood were collected 1 day after challenge from half of the mice in each of the three groups. 

^eVaginal wash for shed virus protein in the vaginal lumen 3 days after challenge. 

^fMice were examined for signs of illness 8 to 14 days after challenge.