Antigen-Experienced CD4lo T Cells Are Linked to Deficient Contraction of the Immune Response in Autoimmune Diabetes

Table 1

The addition of cytokines to the primary stimulation increases the proliferative response in NOD Con A blasts upon secondary challenge with anti-CD3.

Conditions of primary blasts stimulationa	Secondary proliferationb	Stimulation indexc	IFN𝛾 absorbanced
NOD
Con A (no 2^0)	22		0.24
Con A	33∗	1.50∗∗	1.13
Con A+IL-2	30	1.36∗∗	1.01
Con A+IL-4	42∗	1.91	1.21
Con A+IL-7	60∗	2.72∗∗	1.31
Con A+IL-15	90∗	4.09∗∗	1.51
NOR
Con A (no 2^0)	38		0.30
Con A	28∗	0.74	1.54
Con A+IL-2	23∗	0.61	1.35
Con A+IL-4	42	1.11	1.42
Con A+IL-7	42	1.11	1.38
Con A+IL-15	52∗	1.37	1.65

aSplenocytes were cultured for 72 hrs with 2.0 𝜇g/mL of Con A, with or without recombinant mouse IL-2, IL-4, IL-7, or IL-15.
bCon A blasts were plated in triplicate on IgG-coated (no 2⁰) or anti-CD3-coated plates for 48 hrs, pulsed with H³-thymidine for an additional 18 hrs, harvested, and read in a beta counter. The results are the means, rounded to the nearest 10⁴ CPM. The SD was less than 10%. The asterisk (∗) indicates 𝑃<.05, when compared to proliferation on IgG coated plate.
cThe stimulation index (SI) was calculated as mean experimental (anti-CD3 plate) CPM/ mean Con A control (IgG plate) CPM. The two asterisks (∗∗) indicate that 𝑃<.05 when the NOD SI was compared to the SI of NOR blasts treated under the same conditions.
dSupernatants collected 48 hrs after culture on IgG or anti-CD3-coated plates were tested by ELISA. The results are expressed as mean absorbance (405 nm) of duplicate wells. The absorbance values (1.0–1.54) represent an IFN-𝛾 concentration range of 1.5–2.5 ng/mL.