Repeat-Dose Toxicity Study Using the AFPL1-Conjugate Nicotine Vaccine in Male Sprague Dawley Rats
Abstract
:1. Introduction
2. Materials and Methods
2.1. Animals and Husbandry
2.2. Vaccine and Vaccination Protocol
2.3. Clinical Observations and Symptoms
2.4. Euthanasia and Blood Collection
2.5. Hematological and Blood Biochemical Evaluation
2.6. Anatomopathological Studies and Organ Weights
2.7. Immunotoxicological Evaluation
2.8. Anti-Nicotine ELISAs
2.9. Statistical Analysis
3. Results
4. Discussion
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Group | Animals, n | n Euthanized at x Days After 4th Dose | ||
---|---|---|---|---|
x = 3 | x = 7 | x = 14 | ||
PBS | 15 | 5 | 5 | 5 |
Adjuvant (AFPL1) | 15 | 5 | 5 | 5 |
Vaccine | 15 | 5 | 5 | 5 |
Schedule | Route | Volume | ||
Day 0 | IM/IN | 200 µL (100 µL per leg)/80 µL (40 µL per nare) | ||
Day 21 | IN | 80 µL (40 µL per nare) | ||
Day 42 | IN | 80 µL (40 µL per nare) | ||
Day 56 | IN | 80 µL (40 µL per nare) |
Group | Brain | Thymus | Heart | Left Lung | Right Lung | Liver | Spleen | Left Kidney | Right Kidney |
---|---|---|---|---|---|---|---|---|---|
3 Days After 4th Dose | |||||||||
PBS | 0.47 ± 0.01 | 0.10 ±0.01 | 0.32 ± 0.01 | 0.13 ± 0.01 | 0.26 ± 0.01 | 3.01 ± 0.26 | 0.15 ± 0.01 | 0.36 ± 0.01 | 0.36 ± 0.01 |
Adjuvant | 0.47 ± 0.01 | 0.08 ±0.01 | 0.30 ± 0.01 | 0.14 ± 0.01 | 0.28 ± 0.02 | 2.91 ± 0.14 | 0.16 ± 0.01 | 0.35 ± 0.01 | 0.35 ± 0.01 |
Vaccine | 0.48 ± 0.01 | 0.11 ±0.01 | 0.30 ± 0.01 | 0.12 ± 0.01 | 0.24 ± 0.01 | 2.79 ± 0.14 | 0.16 ± 0.01 | 0.35 ± 0.01 | 0.37 ± 0.02 |
7 Days After 4th Dose | |||||||||
PBS | 0.47 ± 0.01 | 0.09 ± 0.01 | 0.35 ± 0.01 | 0.13 ± 0.01 | 0.24 ± 0.01 | 2.96 ± 0.09 | 0.16 ± 0.01 | 0.34 ± 0.01 | 0.34 ± 0.01 |
Adjuvant | 0.49 ± 0.01 | 0.10 ± 0.01 | 0.31 ± 0.01 | 0.13 ± 0.01 | 0.24 ± 0.01 | 2.96 ± 0.20 | 0.17 ± 0.01 | 0.39 ± 0.02 | 0.37 ± 0.02 |
Vaccine | 0.49 ± 0.02 | 0.12 ± 0.03 | 0.33 ± 0.03 | 0.14 ± 0.02 | 0.26 ± 0.03 | 2.79 ± 0.20 | 0.17 ± 0.01 | 0.34 ± 0.01 | 0.36 ± 0.02 |
14 Days After 4th Dose | |||||||||
PBS | 0.49 ± 0.01 | 0.10 ± 0.01 | 0.31 ± 0.01 | 0.13 ± 0.01 | 0.24 ± 0.01 | 3.16 ± 0.11 | 0.17 ± 0.01 | 0.35 ± 0.01 | 0.36 ± 0.02 |
Adjuvant | 0.47 ± 0.02 | 0.10 ± 0.01 | 0.28 ± 0.01 | 0.13 ± 0.01 | 0.24 ± 0.01 | 2.80 ± 0.14 | 0.16 ± 0.01 | 0.36 ± 0.01 | 0.35 ± 0.01 |
Vaccine | 0.44 ± 0.01 | 0.09 ± 0.01 | 0.30 ± 0.01 | 0.12 ± 0.01 | 0.22 ± 0.01 | 3.04 ± 0.10 | 0.17 ± 0.01 | 0.35 ± 0.02 | 0.36 ± 0.02 |
Group | HBG (g/L) | Hto (mL/100L) | LT (103 mm) | PMN (%) | Lf (%) | E (%) | M (%) | Platelets (× 103 µL) |
---|---|---|---|---|---|---|---|---|
3 Days After 4th Dose | ||||||||
PBS | 154.2 ± 4.5 | 51.0 ± 1.6 | 5.2 ± 0.2 | 31.3 ± 1.7 | 68.7 ± 1.7 | 0.0 ± 0.0 | 0.0 ± 0.0 | 746.4 ± 33.0 |
Adjuvant | 154.1 ± 6.3 | 53.3 ± 2.2 | 5.1 ± 0.2 | 33.6 ± 3.8 | 66.1 ± 3.8 | 0.2 ± 0.3 | 0.0 ± 0.0 | 744.2 ± 56.4 |
Vaccine | 157.1 ± 3.6 | 51.5 ± 1.3 | 5.0 ± 0.1 | 30.4 ± 2.3 | 68.4 ± 1.9 | 0.5 ± 0.6 | 0.0 ± 0.0 | 772.3 ± 51.6 |
7 Days After 4th Dose | ||||||||
PBS | 155.0 ± 1.5 | 51.8 ± 0.5 | 5.2 ± 0.1 | 35.6 ± 3.2 | 63.4 ± 3.1 | 0.0 ± 0.0 | 0.0 ± 0.0 | 767.2 ± 43.9 |
Adjuvant | 154.2 ± 2.9 | 51.3 ± 1.0 | 5.1 ± 0.2 | 35.4 ± 2.6 | 63.5 ± 3.3 | 0.1 ± 0.1 | 0.1 ± 0.1 | 797.0 ± 45.3 |
Vaccine | 157.1 ± 3.1 | 53.1 ± 1.5 | 5.2 ± 0.2 | 38.8 ± 3.9 | 58.8 ± 3.4 | 0.1 ± 0.2 | 0.1 ± 0.2 | 790.4 ± 44.5 |
14 Days After 4th Dose | ||||||||
PBS | 157.0 ± 4.1 | 52.4 ± 1.3 | 6.4 ± 0.5 | 34.5 ± 5.1 | 65.3 ± 5.0 | 0.0 ± 0.0 | 0.0 ± 0.0 | 853.1 ± 29.1 |
Adjuvant | 158.1 ± 4.6 | 53.5 ± 2.3 | 5.5 ± 0.2 | 29.3 ± 4.1 | 70.0 ± 3.8 | 0.1 ± 0.4 | 0.0 ± 0.0 | 764.0 ± 23.8 |
Vaccine | 156.0 ± 3.9 | 52.8 ± 1.4 | 6.3 ± 0.5 | 33.7 ± 3.4 | 66.3 ± 3.4 | 0.0 ± 0.0 | 0.0 ± 0.0 | 803.2 ± 30.1 |
(a) | ||||||
Group | ALT (UI) | AST (UI) | TP (g/dL) | Bilirubin (mg/dL) | Urates (µM) | Urea (mM) |
3 Days After 4th Dose | ||||||
PBS | 53.67 ± 9.36 | 167.71 ± 15.38 | 5.11 ± 0.55 | 0.07 ± 0.04 | 64.29 ± 18.31 | 8.15 ± 0.61 |
Adjuvant | 61.02 ± 12.22 | 153.65 ± 12.71 | 5.43 ± 0.42 | 0.16 ± 0.08 | 65.94 ± 17.38 | 8.47 ± 0.91 |
Vaccine | 59.15 ± 10.34 | 159.43 ± 18.49 | 6.10 ± 0.85 | 0.03 ± 0.04 | 57.86 ± 13.56 | 8.27 ± 0.89 |
7 Days After 4th Dose | ||||||
PBS | 74.38 ± 6.44 | 163.45 ± 16.50 | 5.34 ± 0.17 | 0.01 ± 0.04 | 69.90 ± 10.55 | 9.47 ± 1.19 |
Adjuvant | 62.71 ± 9.53 | 154.88 ± 14.97 | 5.47 ± 0.40 | 0.06 ± 0.03 | 79.13 ± 18.29 | 8.82 ± 1.23 |
Vaccine | 70.13 ± 11.19 | 161.26 ± 14.30 | 5.11 ± 0.38 | 0.08 ± 0.04 | 66.31 ± 7.73 | 8.64 ± 0.93 |
14 Days After 4th Dose | ||||||
PBS | 56.45 ± 7.93 | 128.33 ± 11.69 | 5.92 ± 0.22 | 0.01 ± 0.01 | 87.92 ± 17.11 | 8.14 ± 1.50 |
Adjuvant | 65.51 ± 13.70 | 133.53 ± 13.32 | 5.94 ± 0.41 | 0.10 ± 0.04 | 66.27 ± 11.23 | 8.95 ± 1.25 |
Vaccine | 42.32 ± 6.37 | 152.44 ± 18.57 | 6.82 ± 0.77 | 0.09 ± 0.04 | 68.32 ± 11.35 | 9.70 ± 1.03 |
(b) | ||||||
Group | CPK (UI) | Creatinine (µM) | Glucose (mM) | ALP (UI) | Triglycerides (mM) | Cholesterol (mM) |
3 Days After 4th Dose | ||||||
PBS | 1082.93 ± 201.29 | 65.12 ± 2.87 | 7.45 ± 0.52 | 282.82 ± 17.32 | 0.39 ± 0.12 | 1.25 ± 0.18 |
Adjuvant | 1229.24 ± 153.11 | 68.69 ± 6.18 | 8.39 ± 0.67 | 288.32 ± 30.09 | 0.76 ± 0.24 | 1.72 ± 0.26 |
Vaccine | 1189.94 ± 80.87 | 63.16 ± 8.77 | 9.21 ± 0.80 | 287.24 ± 16.14 | 0.94 ± 0.20 | 1.29 ± 0.10 |
7 Days After 4th Dose | ||||||
PBS | 1488.94 ± 173.82 | 60.83 ± 4.61 | 9.70 ± 1.70 | 410.41 ± 38.78 | 0.85 ± 0.19 | 1.18 ± 0.19 |
Adjuvant | 1362.28 ± 91.59 | 61.32 ± 4.97 | 8.85 ± 1.28 | 452.10 ± 57.35 | 0.59 ± 0.14 | 1.25 ± 0.16 |
Vaccine | 1228.22 ± 49.77 | 56.60 ± 3.85 | 10.18 ± 1.22 | 434.26 ± 33.24 | 0.63 ± 0.17 | 0.99 ± 0.13 |
14 Days After 4th Dose | ||||||
PBS | 1334.75 ± 150.25 | 75.83 ± 8.27 | 10.47 ± 0.89 | 289.93 ± 20.36 | 1.17 ± 0.22 | 1.27 ± 0.11 |
Adjuvant | 1302.22 ± 120.83 | 78.14 ± 11.44 | 9.46 ± 1.14 | 264.22 ± 43.21 | 1.07 ± 0.30 | 1.30 ± 0.18 |
Vaccine | 1422.27 ± 134.45 | 66.49 ± 7.49 | 10.42 ± 1.12 | 213.89 ± 22.83 | 1.51 ± 0.32 | 1.14 ± 0.09 |
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Share and Cite
Oliva, R.; Fraleigh, N.L.; Lewicky, J.D.; Fariñas, M.; Hernández, T.; Martel, A.L.; Navarro, I.; Dagmar, G.-R.; Acevedo, R.; Le, H.-T. Repeat-Dose Toxicity Study Using the AFPL1-Conjugate Nicotine Vaccine in Male Sprague Dawley Rats. Pharmaceutics 2019, 11, 626. https://doi.org/10.3390/pharmaceutics11120626
Oliva R, Fraleigh NL, Lewicky JD, Fariñas M, Hernández T, Martel AL, Navarro I, Dagmar G-R, Acevedo R, Le H-T. Repeat-Dose Toxicity Study Using the AFPL1-Conjugate Nicotine Vaccine in Male Sprague Dawley Rats. Pharmaceutics. 2019; 11(12):626. https://doi.org/10.3390/pharmaceutics11120626
Chicago/Turabian StyleOliva, Reynaldo, Nya L. Fraleigh, Jordan D. Lewicky, Mildrey Fariñas, Tamara Hernández, Alexandrine L. Martel, Ingrid Navarro, García-Rivera Dagmar, Reinaldo Acevedo, and Hoang-Thanh Le. 2019. "Repeat-Dose Toxicity Study Using the AFPL1-Conjugate Nicotine Vaccine in Male Sprague Dawley Rats" Pharmaceutics 11, no. 12: 626. https://doi.org/10.3390/pharmaceutics11120626
APA StyleOliva, R., Fraleigh, N. L., Lewicky, J. D., Fariñas, M., Hernández, T., Martel, A. L., Navarro, I., Dagmar, G.-R., Acevedo, R., & Le, H.-T. (2019). Repeat-Dose Toxicity Study Using the AFPL1-Conjugate Nicotine Vaccine in Male Sprague Dawley Rats. Pharmaceutics, 11(12), 626. https://doi.org/10.3390/pharmaceutics11120626