Molecular Biology Reports

, Volume 39, Issue 3, pp 3225–3233 | Cite as

In vivo study of hepatitis B vaccine effects on inflammation and metabolism gene expression

  • Heyam Hamza
  • Jianhua Cao
  • Xinyun Li
  • Shuhong ZhaoEmail author


Pharmaceutical companies usually perform safety testing of vaccines, but all requirements of the World Health Organization and drug pharmacopoeias depend on general toxicity testing, and the gene expression study of hepatitis B vaccine is not done routinely to test vaccine quality. In this study, we applied a new technique of gene expression analysis to detect the inflammation and metabolism genes that might be affected by hepatitis B vaccine in mouse liver. Mice were used and divided into three groups: the first and second groups were treated with one or two human doses of vaccine, respectively, and the third group was used as a control. A microarray test showed that expression of 144 genes in the liver was significantly changed after 1 day of vaccination. Seven of these genes, which were related to inflammation and metabolism, were chosen and confirmed by quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) at 1, 4 and 7 days. The expression level of these genes can be considered as a biomarker for the effects of the vaccine.


Hepatitis B vaccine Inflammation genes Metabolism genes DNA microarray Quantitative RT-PCR analysis 



This research was supported by key project of National Natural Science Foundation of China (U0631005). We thank Yang Feng, GU Ting, Wong Feng Li and Shen He in our lab for sample collections and suggestions on data analyses.

Supplementary material

11033_2011_1090_MOESM1_ESM.doc (96 kb)
Supplementary material 1 (DOC 96 kb)


  1. 1.
    Kurata Takeshi (2006) Minimum requirements for biological products. National institute of infectious diseases, Japan, p 130Google Scholar
  2. 2.
    Martinuc Porobic J, Avcin T, Bozic B, Kuhar M, Cucnick S, Zupancic M, Prosenc K, Kveder T, Rozman B (2005) Anti-phospholipids antibodies following Vaccination with recombinant hepatitis vaccine. J Clin Exp Immunol 142:377–380CrossRefGoogle Scholar
  3. 3.
    Maillefert JF, Sibilia J, Toussirot E, Vignon E, Eschard JP, Lorcerie B, Gaudin R (1999) Rheumatic disorders developed after hepatitis B vaccination. Rheumatology 38:978–983PubMedCrossRefGoogle Scholar
  4. 4.
    Good PF, Perl DP, Bierer LM, Schmeidler J (1992) Selective accumulation of aluminum and iron in the neurofibrillary tangles of Alzheimer’s disease: a laser microprobe (LAMMA) study. Ann Neurol 31(3):286–292PubMedCrossRefGoogle Scholar
  5. 5.
    Flarend RE, Hem SL, White JL, Elmore D, Suchow MA, Rudy AC, Dandashli EA (1997) In vivo absorption of aluminum-containing vaccine adjuvant using26Al. Vaccine 15:1314–1318PubMedCrossRefGoogle Scholar
  6. 6.
    British Pharmacopoeia Commission (2003) Test for abnormal toxicity. (Ph.Eur.method 2.6.9) Volume IV Appendices Appendix XIV E General test 1, TSO publisherGoogle Scholar
  7. 7.
    Milstien J, Grachev V, Padilla A, Griffiths E (1996) WHO activities towards the three Rs in the development and control of biological products. Dev Biol Stand 86:31–39PubMedGoogle Scholar
  8. 8.
    Mizukami T, Masumi A, Momose H, Kuramitsu M, Takizawa K, Naito S, Maeyama J-I, Furuhata K, Tsuruhara M, Hamaguchi I, Yamaguchi K (2009) An improved abnormal toxicity test by using reference vaccine-specific body weight curves and histopathological data for monitoring vaccine quality and safety in Japan. Biological 37(1):8–17CrossRefGoogle Scholar
  9. 9.
    Hamaguchi I, Imai J, Momose H, Kawamura M, Mizukami T, Naito S, Maeyama J, Masumi A, Kuramitsu M, Takizawa K, Kato H, Mizutani T, Horiuchi Y, Nomura N, Watanabe S, Yamaguchi K (2008) Application of quantitative gene expression analysis for pertussis vaccine safety control. Vaccine 26:4686–4696PubMedCrossRefGoogle Scholar
  10. 10.
    Hamaguchi I, Imai J, Momose H, Kawamura M, Mizukami T, Kato H, Naito S, Maeyama J, Masumi A, Kuramitsu M, Takizawa K, Mochizuki M, Ochiai M, Yamamoto A, Horiuchi Y, Nomura N, Watanabe S, Yamaguchi K (2007) Two vaccine toxicity-related genes Agp and Hpx could prove useful for pertussis vaccine safety control. Vaccine 25(17):3355–3364PubMedCrossRefGoogle Scholar
  11. 11.
    Fengmei H, Ming C, Qisong X, Yong C (2006) Gene expression profile in immunologically injured liver cell of mice. Sci China Series C: Life Sci 49(5):454–459CrossRefGoogle Scholar
  12. 12.
    Mizukami T, Imai J, Hamaguchi I, Kawamura M, Momose H, Naito S, Maeyama J, Masum A, Kuramitsu M, Takizawa K, Nomura N, Watanabe S, Yamaguchi K (2008) Application of DNA microarray technology to influenza A/Vietnam/1194/2004 (H5N1) vaccine safety evaluation. Vaccine 26(4):2270–2283PubMedCrossRefGoogle Scholar
  13. 13.
    Irizarry RA, Bolstad BM, Collin F, Cope LM, Hobbs B, Speed TP (2003) Summaries of Affymetrix GeneChip probe level data. Nucleic Acids Res 31(4):1–8CrossRefGoogle Scholar
  14. 14.
    Dai X, Xiang O, Hao X, Cao D, Tang Y, Yanwei H, Xiaoxu L, Tang C (2007) Effect of T0901317 on hepatic proinflammatory gene expression in ApoE/mice fed a high-fat/high-cholesterol diet. Inflammation 30(8):105–117PubMedCrossRefGoogle Scholar
  15. 15.
    Momose H, Imai J, Hamaguchi I, Kawamura M, Mizukami T, Naito S, Masumi A, Maeyama J, Takizawa K, Kuramitsu M, Nomura N, Watanabe S, Yamaguchi K (2010) Induction of indistinguishable gene expression pattern in Rats by Vero-cell derived and mouse-brain derived Japanese encephalitis vaccines. Japan J Infect Dis 63:25–30Google Scholar
  16. 16.
    Alex Merrick B (2006) Toxicoproteomics in liver injury and inflammation. Ann NY Acad Sci 1076:707–717PubMedCrossRefGoogle Scholar
  17. 17.
    Gutfeld O, Prus D, Ackerman Z, Dishon S, Linke RP, Levin M, Urieli-Shoval S (2006) Expression of serum amyloid a, in normal, dysplastic, and neoplastic human colonic mucosa: implication for a role in colonic tumorigenesis. J Histochem Cytochem Jan 54(1):63–73CrossRefGoogle Scholar
  18. 18.
    Liu Z, Ukomadu C (2008) Fibrinogen-like-protein 1, a hepatocyte derived protein is an acute phase reactant. Biochem Biophys Res Commun 25(1):729–773CrossRefGoogle Scholar
  19. 19.
    Marhaug G, Bruce Dowton S (1994) Serum amyloid A: an acute phase apolipoprotein and precursor of AA amyloid. Baillieres Clin Rheumatol 8(3):553–573PubMedCrossRefGoogle Scholar
  20. 20.
    Stríz I, Trebichavský I (2004) Calprotectin: a pleiotropic molecule in acute a cute and chronic inflammation. Physiol Res 53(3):245–253PubMedGoogle Scholar
  21. 21.
    Gonzalez MA, Alvarez Mdel L, Pisani GB, Bernal CA, Roma MG, Carrillo MC (2007) Involvement of oxidative stress in the impairment in biliary secretory function induced by intraperitoneal administration of aluminum to rats. Biol Trace Elem Res 116(3):329–348. doi: 10.1007/BF02698017 PubMedCrossRefGoogle Scholar
  22. 22.
    Momose H, Mizukami T, Ochiai M, Hamaguchi I, Yamaguchi K (2010) A new method for the evaluation of vaccine safety based on comprehensive gene expression analysis. J Biomed Biotechnol 2010:361841:1–7. doi: 10.1155/2010/361841 Google Scholar
  23. 23.
    Kindy MS, de Beer MC, Jin Y, de Beer FC (2000) Expression of mouse acute phase (SAA1.1) and constitutive (SAA4) serum amyloid a isotypes: influence on lipoprotein profiles. Arterioscler Thromb Vasc Biol 20:1543–1550PubMedCrossRefGoogle Scholar
  24. 24.
    Khovidhunkit W, Kim M-S, Memon RA, Shigenaga JK, Moser AH, Fenigold KR, Grunfeld C (2004) Effects of infection and Inflammation on lipid and lipoprotein metabolism: mechanisms and consequences to the host. J Lipid Res 45:1169–1196PubMedCrossRefGoogle Scholar
  25. 25.
    Ribeiro FP, Furlaneto CJ, Hatanaka E, Ribeiro WB, Souza GM, Cassatella MA, Campa A (2003) mRNA expression and release of interleukin-8 induced by serum amyloid A in neutrophils and monocytes. Mediators Inflam 12(3):173–178CrossRefGoogle Scholar
  26. 26.
    Chait A, Han CY, Oram JF, Heinecke JW (2005) Lipoprotein-associated inflammatory proteins markers or mediators cardiovascular disease. J Lipid Res 46:389–403PubMedCrossRefGoogle Scholar
  27. 27.
    Sullivan CP, Seidl SE, Rich CB, Raymondjean M, Schreiber BM (2010) Secretory phospholipase A2, group IIA is a novel serum amyloid a target gene. J Biol Chem 285(1):565–575PubMedCrossRefGoogle Scholar
  28. 28.
    Passey RJ, William E, Lichanska AM, Wells C, Hu S, Geczy CL et al (1999) A null mutation in the inflammation-associated S100 protein S100A8 causes early resorption of the mouse embryo. J Immunol 163:2209–2216PubMedGoogle Scholar
  29. 29.
    Ebihara T, Endo R, Kikuta H, Ishiguro N, Ma X, Shimazu M, Otoguro T, Kobayashi K (2005) Differential gene expression of S100 protein family in leukocytes from patients with Kawasaki disease. Eur J Pediatr 164:427–431PubMedCrossRefGoogle Scholar
  30. 30.
    Yui S, Nakatani Y, Mikami M (2003) Calprotectin (S100A8/S100A9), an inflammatory protein Complex from neutrophils with a broad apoptosis-inducing activity. Biol Pharm Bull 26(6):753–756PubMedCrossRefGoogle Scholar
  31. 31.
    Rosen MB, Thibodeaux JR, Wood CR, Zehr RD, Schmid JE, Lau C (2007) Gene expression profiling in the lung and liver of PFOA-exposed mouse fetuses. Toxicology 239(1–2):15–33PubMedCrossRefGoogle Scholar
  32. 32.
    Cadoudal T, Blouin JM, Collinet M, Fouque F, Tan GD, Loizon E, Beale G, Frayn KN, Karpe F, Vidal H, Benelli H, Forest C (2007) Acute and selective regulation of glyceroneogenesis and cytosolic phosphoenolpyruvate carboxykinase in adipose tissue by thiazolidinediones in type 2 diabetes. Diabetology 50:666–675CrossRefGoogle Scholar
  33. 33.
    Nichols KD, Kirby GM (2008) Microarray analysis of hepatic gene expression in pyrazole-mediated hepatotoxicity: identification of potential stimuli of Cyp2a5. Biochem Pharmacol 75:538–551PubMedCrossRefGoogle Scholar
  34. 34.
    Frijters R, Fleuren W, Toonen EJM, Tuckermann JP, Reichardt HM, van der Maaden H, van Elsas A, van Lierop M-J, Dokter W, de Vlieg J, Alkema W (2010) Research article prednisolone-induced differential gene expression in mouse liver carrying wild type or a dimerization-defective glucocorticoid receptor. BMC Genomics 11(359):1–14Google Scholar
  35. 35.
    Beal EG, Harveand BJ, Forest C (2007) PCK1 and PCK2 as candidate diabetes and obesity genes. Cell Biochem Biophys 48(2–3):89–95. doi: 10.1007/s12013-007-0025-6 CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  • Heyam Hamza
    • 1
  • Jianhua Cao
    • 1
  • Xinyun Li
    • 1
  • Shuhong Zhao
    • 1
    Email author
  1. 1.Key Lab of Animal Genetics, Breeding, and Reproduction of Ministry of Education, College of Animal Science and TechnologyHuazhong Agricultural UniversityWuhanPeople’s Republic of China

Personalised recommendations