Table 1.
Young Adult Women.
Table 2.
Post-menopausal (P.M.) Women no HT.
Table 3.
Post-menopausal (P.M.) Women receiving ET.
Fig 1.
Impact of menopause and ET on lymphocyte frequencies.
(A) The number of lymphocytes on visit 1 /μl blood based on complete blood cell counts (CBC) values. (B) Frequency of CD4 T cells, CD8 T cells and CD20 B cells was determined in PBMC using flow cytometry (FCM). The percentages were then converted to absolute numbers of cells/μl blood using the lymphocyte counts obtained from the CBCs. (C) Frequency of naïve (Na), central memory (CM) and effector memory (EM) CD4 T cells was determined using FCM and were then converted to number/μl of blood using the CD4 numbers obtained earlier. (D) Numbers of Na, CM and EM CD8 T cells was determined as described for CD4 T cells. (E) Frequencies of naïve (CD27-) and memory (CD27+) B cells were determined by FCM and then converted to absolute numbers of cells/μl blood using the CD20 numbers obtained earlier.
Fig 2.
Impact of menopause and ET on IL-6 levels and TNFα and IFNγ production.
(A) Frequency of CD4 and CD8 T cells that secrete TNFα, IFNγ or both in response to CD3 stimulation was determined by intracellular cytokine staining and FCM using PBMC collected during visit 1 before the administration of the influenza vaccine. (B) Plasma IL-6 levels using samples collected during visit 1 were determined by ELISA.
Fig 3.
Impact of menopause and ET on immune response to seasonal influenza vaccine.
(A) IgG end point titers were determined using standard ELISA and then the fold increase over baseline was calculated for visits 2 and 3. (B) Correlation between plasma E2 levels on visit 1 (day of vaccination) and fold increase in IgG titer during visit 2 was calculated for women in group C. (C) Frequency of flu-specific CD4 T cells was measured using intracellular cytokine staining and flow cytometry. The increased frequency of responding T cells was determined by subtracting the values obtained on visits 2 and 3 from those obtained on visit 1. (D) Same analysis was carried out for CD8 T cells.