Table II.

Chlamydia pneumoniae peptide-stimulated effector cells: cytokine and cell surface phenotype profiles

IDPeptideIFN-γ (ng/ml)aIL-4 (pg/ml)aTNF-α (pg/ml)aTCRαβ+CD8+CD44+b
Stimulation cycleStimulation cycleStimulation cycleStimulation cycle
121223 (%)
Cpn CTL peptides
2QESCYGRL2.518.5<8<8243.893.7
3ISVSNPGDL1.910.8<8<8198.493.1
13STEINQPFITM12.561.2<8<8597.198.9
17YASDNQAIL3.015.4<8<8206.396.8
19LVYNYPGV5.124.6<8<8281.097.0
20LIYNYPGV4.324.8<8<8275.297.7
22LIFNYPGV2.811.5<8<8211.990.0
23HPYLYRLL3.013.9<8<8187.790.2
24HPTLFKVL2.818.1<8<8266.593.3
25SIILFLPL6.139.9<8<8494.195.9
27ISNGNSDCL3.226.1<8<8289.994.1
30QAPTNRWML5.230.3<8<8328.695.8
31SLLGNATAL1.913.3<8<8185.390.7
32SHYAFSPMFEVL2.919.9<8<8292.094.0
33ISFAFCQL3.230.1<8<8338.792.8
36GTYHFTKL4.922.6<8<8303.897.6
37FQLCNSYDL5.649.5<8<8386.798.9
39LQQRYSRL12.857.9<8<8504.699.2
Negative Cpn peptides
11TAGANPMDL0.41.1<8<867.215.1
12ISANNDSEI0.050.6<8<855.410.6
16LMSGFRQM<0.040.6<8<8<40.011.7
21LLVFNYPGI0.91.6<8<862.514.9
TcTSAVDYNFTIV5.338.4<8<8354.396.2
  • a Levels of IFN-γ, IL-4, and TNF-α were determined by ELISA as described in Materials and Methods. Culture supernatants were harvested 48 h poststimulation with peptide (2 μM) and kept at −20°C until assayed. Negative and positive control supernatants were obtained from Cpn- and T. cruzi-infected mice using SC stimulated with Cpn peptides with no CTL activity and TcTSA, respectively. Supernatants from wells containing Cpn CTL peptide-stimulated naive SC contained minimal to undetectable levels of cytokines.

  • b Effector cells purified at the end of the third cycle of stimulation with Cpn CTL peptides were analyzed by flow cytometry for surface expression of CD3, TCRαβ, CD8, CD4, and CD44 after staining with FITC- and PE-conjugated mAbs. Negative and positive control effector cell lines were SC from Cpn- and T. cruzi-infected mice stimulated for three cycles with Cpn peptides negative for CTL activity and TcTSA, respectively. Similar cytokine levels and cell surface phenotypes were obtained in two to four additional experiments.