Advancing Prevention of STIs by Developing Specific Serodiagnostic Targets: Trichomonas vginalis as a Model
Abstract
:1. Introduction
2. Materials and Methods
2.1. Epitopes Unique to the T. vaginalis A, E and G Proteins
2.2. ACT-P2, a Truncated α-Actinin Protein Used for Screening of Patient Sera
2.3. Plasmid Encoding AEG::SOE2 and ACT-P2
2.4. Recombinant Proteins
2.5. Sera for Enzyme-Linked Immunosorbent Assay (ELISA) and Derivation of Positive/Negative (P/N) Scores
2.6. Mouse Anti-AEG::SOE2 Serum and Anti-T. vaginalis Serum and ELISA for Detecting Antibody to T. vaginalis
2.7. Reproducibility
3. Results
3.1. Epitopes of A, E, and G Unique to T. vaginalis and the AEG::SOE2 Protein Sequence
3.2. Purification of Recombinant AEG::SOE2
3.3. ELISA Using Different Amounts of AEG::SOE2-Coated Wells Probed with Different Antibodies
3.4. ELISA Comparing ACT-P2 and AEG::SOE2 with Different Antibodies
3.5. Different ELISA Experiments Testing Women and Men Sera of Different 0 to 4+ Scores
3.6. ELISA of Women and Men Sera of 3+ and 4+ Scores at Different Dilutions
3.7. ELISA Using ACT-P2 and AEG::SOE2 as Targets with 0 to 4+ Individual Sera
3.8. AEG::SOE2 is Immunogenic, and Anti-AEG::SOE2 Serum IgG Antibody Detects T. vaginalis Organisms Immobilized onto Microtiter Wells by ELISA
4. Discussion
5. Conclusions
6. Patent
Funding
Acknowledgments
Conflicts of Interest
References
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Proteins | |||
---|---|---|---|
No. | A | E | G |
1 | HTYTRPEEVODFVSK | AEHDAIVKECIAEEAA | RACRKLYPKDIQVVA |
2 | LGEARTKLTEMYMRK | VKYLGRVTLAARSSA | NVYLLKYDTAHRAFP |
3 | VTLAARSSAPSGAST | QEFTVGEGADKWVVK | |
4 | TDGTVLKKNIGGNAC | WVVKSIGGRLGPSQL | |
5 | DKVPKKFKLPSPFFN | VVLESTGIFRTKAEK | |
6 | KLGGLLVKKYGLSAK | KDAEGKIKKDDGYDGH | |
7 | ILVKKYGLSAKNLDEF | TLNNAFGIRNGFMTTV | |
8 | ASSEFYDEEKKLYEV | ||
9 | DYENWTKLNARLGQRV |
Expt | Serum Reaction Scores Against ACTP2: | |||||||||||||
0‡‡ | 0 | 1+ | 1+ | 2+ | 2+ | 3+ | 3+ | 4+ | 4+ | 4+ | 4+ | 4+ | 4+ | |
W | M | W | M | W | M | W | M | W | M | W | M | W | M | |
expt 1 | 0 | 0 | 1 | 1 | 2 | 2 | 3 | 3 | 4 | 4 | 4 | 4 | 4 | 4 |
expt 2 | 0 | 0 | 1 | 1 | 2 | 2 | 3 | 3 | 4 | 4 | 4 | 4 | 4 | 4 |
expt 3 | 0 | 0 | 1 | 1 | 2 | 2 | 3 | 3 | 4 | 4 | 4 | 4 | 4 | 4 |
expt 4 | 0 | 0 | 1 | 1 | 2 | 2 | 3 | 3 | 4 | 4 | 4 | 4 | 4 | 4 |
Expt | Serum Reaction Scores Against AEG::SOE2: | |||||||||||||
0 | 0 | 1+ | 1+ | 2+ | 2+ | 3+ | 3+ | 4+ | 4+ | 4+ | 4+ | 4+ | 4+ | |
W | M | W | M | W | M | W | M | W | M | W | M | W | M | |
expt 1 | 0 | 0 | 1 | 1 | 2 | 2 | 3 | 3 | 4 | 4 | 4 | 4 | 4 | 4 |
expt 2 | 0 | 0 | 1 | 1 | 2 | 2 | 3 | 3 | 4 | 4 | 4 | 4 | 4 | 4 |
expt 3 | 0 | 0 | 1 | 1 | 2 | 2 | 3 | 3 | 4 | 4 | 4 | 4 | 4 | 4 |
expt 4 | 0 | 0 | 1 | 1 | 2 | 2 | 3 | 3 | 4 | 4 | 4 | 4 | 4 | 4 |
No.‡ | Scores # | Scores | No. | Scores | Scores | No. | Scores | Scores | No. | Scores | Scores |
---|---|---|---|---|---|---|---|---|---|---|---|
AEG::SOE2 | ACTP2 | AEG::SOE2 | ACTP2 | AEG::SOE2 | ACTP2 | AEG::SOE2 | ACTP2 | ||||
1 | 0 | 0 | 22 | 4+ | 3++ | 43 | 4+ | 4+ | 64 | 3+ | 2 |
2 | 4+ | 4+ | 23 | 3+ | 3+ | 44 | 3+ | 3+ | 65 | 4+ | 3++ |
3 | 3+ | 2++ | 24 | 4+ | 4+ | 45 | 3+ | 3+ | 66 | 2+ | 2+ |
4 | 1+ | 2+ | 25 | 4+ | 4+ | 46 | 4+ | 3+ | 67 | 3+ | 3++ |
5 | 2+ | 3+ | 26 | 3++ | 3+ | 47 | 2+ | 2+ | 68 | 2+ | 2+ |
6 | 2+ | 2+ | 27 | 4+ | 3++ | 48 | 3++ | 3+ | 69 | 2+ | 2+ |
7 | 3+ | 4+ | 28 | 4+ | 3++ | 49 | 3+ | 3+ | 70 | 3+ | 3++ |
8 | 2+ | 2+ | 29 | 2+ | 2+ | 50 | 2+ | 3+ | 71 | 1+ | 1+ |
9 | 2+ | 2+ | 30 | 1+ | 1+ | 51 | 3+ | 2+ | 72 | 0 | 0 |
10 | 4+ | 4+ | 31 | 1+ | 1+ | 52 | 4+ | 3++ | 73 | 1+ | 1+ |
11 | 4+ | 3++ | 32 | 1+ | 2+ | 53 | 3+ | 2+ | 74 | 0 | 0 |
12 | 2+ | 3+ | 33 | 4+ | 3++ | 54 | 3+ | 1+ | 75 | 2+ | 3+ |
13 | 1+ | 34 | 4+ | 3+ | 55 | 3++ | 3+ | 76 | 0 | 1+ | |
14 | 4+ | 4+ | 35 | 3++ | 4+ | 56 | 2+ | 2+ | 77 | 4+ | 4+ |
15 | 0 | 0 | 36 | 3+ | 2+ | 57 | 3+ | 3++ | 78 | 3++ | 3++ |
16 | 2+ | 2+ | 37 | 3+ | 3+ | 58 | 4+ | 4+ | 79 | 3+ | 3+ |
17 | 4+ | 4+ | 38 | 3+ | 2+ | 59 | 3+ | 3+ | 80 | 3+ | 3+ |
18 | 3+ | 3+ | 39 | 3++ | 3+ | 60 | 4+ | 4+ | 81 | 4+ | 4+ |
19 | 4+ | 3++ | 40 | 1+ | 1+ | 61 | 4+ | 3++ | 82 | 3+ | 3+ |
20 | 3++ | 4+ | 41 | 2+ | 1+ | 62 | 3+ | 3+ | 83 | 4+ | 4+ |
21 | 4+ | 4+ | 42 | 3+ | 3+ | 63 | 3+ | 3+ | 84 | 4+ | 4+ |
Experiment No. (Mean ± SD) | |||
---|---|---|---|
Antibodies * | 1 | 2 | 3 |
goat anti-mouse IgG alone | 0.075 ± 0.001 ‡ | 0.123 ± 0.005 | 0.130 ± 0.015 |
NMS (1:10) | 0.160 ± 0.001 | 0.157 ± 0.002 | 0.140 ± 0.010 |
anti-T. vaginalis serum (1:10) | ND ‡‡ | 0.364 ± 0.005 | 0.455 ± 0.005 |
anti-T. vaginalis serum (1:100) | 0.345 ± 0.005 | ND | 0.432 ± 0.001 |
anti-AEG::SOE2 serum (1:10) | 0.225 ± 0.002 | ND | ND |
anti-AEG::SOE2 serum (1:10) | 0.207 ±0.002 | 0.305 ± 0.005 | 0.190 ± 0.005 |
MAb cocktail (1:1:1:1, v/v) | 0.275 ± 0.005 | 0.218 ± 0.005 | 0.228 ± 0.005 |
MAb ALD30A | 0.335 ± 0.010 | 0.220 ± 0.010 | 0.253 ± 0.008 |
MAb L64 | 0.055, 0.055 | 0.051, 0.052 | 0.051, 0.050 |
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Alderete, J.F. Advancing Prevention of STIs by Developing Specific Serodiagnostic Targets: Trichomonas vginalis as a Model. Int. J. Environ. Res. Public Health 2020, 17, 5783. https://doi.org/10.3390/ijerph17165783
Alderete JF. Advancing Prevention of STIs by Developing Specific Serodiagnostic Targets: Trichomonas vginalis as a Model. International Journal of Environmental Research and Public Health. 2020; 17(16):5783. https://doi.org/10.3390/ijerph17165783
Chicago/Turabian StyleAlderete, John F. 2020. "Advancing Prevention of STIs by Developing Specific Serodiagnostic Targets: Trichomonas vginalis as a Model" International Journal of Environmental Research and Public Health 17, no. 16: 5783. https://doi.org/10.3390/ijerph17165783