Journal of Diabetes Research

Research Article

Application of IgG-Derived Natural Treg Epitopes (IgG Tregitopes) to Antigen-Specific Tolerance Induction in a Murine Model of Type 1 Diabetes

Table 3

Two-step assay results. Diabetic and healthy subject PBMCs were incubated for 7 days with a pool of GAD65 epitopes with or without Tregitopes. The cells were then washed and put into ELISpot plates to test recall responses to individual epitopes, the same pool used in the primary culture, or whole GAD65 protein. ELISpot results were considered positive if the number of SFC over background per million cells was >50 (Bold). Spot counts that failed to meet this threshold were considered negative results (Italic). Due to variable PBMC yield and recovery after 7 days, not all epitopes were included in every ELISpot, as indicated by “ND”.
 Subject D1100HSubject D1101HSubject D1104HSubject D1105HSubject D1106HSubject D1107H
 TregitopeTregitopeTregitopeTregitopeTregitopeTregitope
 −+−+−+−+−+−+
GAD65_450–470−48 −48 26032477697319 * 15524 * 22 −18*
GAD65_550–570199141126100146101352228 * 881131601 **
GAD65_504–518101−66*31 −12 7211 7728 82747 −18
GAD65_480–4965139 7022 * 12941 * 754708842 −3
GAD65_511–525186−44*3632 6 26 2 −4 5423 18 −13
GAD65_264–2879133 6126 * 141154205138 * 17366 * 48−4*
GAD65_369–39211211351254283434266 ** 468406NDNDNDND
GAD65_355–37220551511949 * 151268488509NDNDNDND
GAD65_203–226NDND15751221154245138 * 207138NDND
GAD65_243–267NDND14635 ** 715430 274134144NDND
GAD65_405–424NDND715612664NDND324365NDND
GAD65_086–1031491066938 21 1 −32 765946 13 −19
GAD65_338–354NDND7644 54−4*NDND48 24 NDND
GAD65_175–190NDND40 23 7259NDND33 17 NDND
GAD65 protein4137737 5932 69NDND6705919546
GAD65 peptide pool168114294032593913786455795525432629 **
 Negative Control 669Negative Control 668Negative Control 674Negative Control 670Negative Control 672  
 TregitopeTregitopeTregitopeTregitopeTregitope  
 −+−+−+−+−+  
GAD65_450–47033 13 125−10**39646411523 * −98 63  
GAD65_550–57014430 ** 31244049043125 −90 NDND  
GAD65_504–51824 27 22275 * 21 7639 −26 0 133  
GAD65_480–49631398 ** 19519316269 ** −28 −81 8 49   
GAD65_511–52513 −17*13810319 24 36 −79 52113  
GAD65_264–28726 12 81359318315312919 5 39   
GAD65_369–392NDNDNDND750531 * NDNDNDND  
GAD65_355–372NDNDNDND7217 * NDNDNDND  
GAD65_203–2261095247546016810932982 ** NDND  
GAD65_243–26721 −12 287207 * 161134188−9**6759  
GAD65_405–4247813 ** 178−8**16 26 49 −14 −10 11   
GAD65_086–10311 5 1431285134 27 −9 −27 16   
GAD65_338–354NDNDNDND4 14 NDNDNDND  
GAD65_175–19039 −3 NDND11 8 53−7*NDND  
GAD65 proteinNDNDNDND44 818NDNDNDND  
GAD65 peptide pool7630 82069261342 117−19 10048   
50 SFC over BG/106.
50 SFC over BG/ .
* .
** .