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CD40mAb adjuvant induces a rapid antibody response that may be beneficial in post-exposure prophylaxis
Journal of Immune Based Therapies and Vaccines volume 8, Article number: 1 (2010)
Active vaccination can be effective as a post-exposure prophylaxis, but the rapidity of the immune response induced, relative to the incubation time of the pathogen, is critical. We show here that CD40mAb conjugated to antigen induces a more rapid specific antibody response than currently used immunological adjuvants, alum and monophosphoryl lipid A™.
Post-exposure prophylaxis (PEP), or the induction of protection against an infectious disease after exposure to the pathogen, is either utilised or has been suggested as an appropriate course of action for a number of diseases, including rabies , anthrax , varicella [3, 4], HIV and Hepatitis A . PEP can be divided into three categories: The administration of antibiotics or antivirals, passive immunization using specific immunoglobulin, and active immunization (vaccination).
In some cases appropriate antimicrobial chemotherapy may not be available, or there may be a worry that the pathogen could be resistant to the agent, particularly in bioterrorism cases. Passive immunization using immunoglobulin may be a suitable alternative, if the pathogen is susceptible to antibody-mediated killing. However, active vaccination has the potential advantages of lower cost, less risk of adventitious pathogen transfer, and most importantly the induction of long-term protection. The use of active vaccination as PEP however depends upon the rapidity with which a protective immune response can be generated, in comparison with the incubation period of the pathogen post-exposure. The kinetics of the immune response are therefore a potential rate-limiting step for the efficacy of post-exposure vaccination.
Immune responses against vaccines are enhanced by immunological adjuvants. Aluminum salts are the only widely licensed immunological adjuvants , but the adjuvant monophosphoryl lipid A (MPL™) is now licensed in some countries for use in the cervical cancer vaccine, Cervarix™  and may shortly be licensed for use in a wider range of vaccines. Aside from MPL™, there is a large amount of research ongoing into other potential adjuvants, including host co-stimulatory molecules , TLR agonists [6, 9], other particulate carriers [10, 11] and combinations of these approaches [12, 13].
Agonistic antibodies against the antigen presenting cell surface antigen CD40 are able to mimic the effect of binding of the ligand, CD154, both in vitro  and in vivo . We have shown that agonistic CD40monoclonal antibody (mAb) is an effective immunological adjuvant at low doses when chemically conjugated to antigen. It is able to enhance antibody [16, 17] as well as T helper responses . CD40 antibody or ligand is also being investigated in cancer therapy and vaccination [19, 20]. As we believe CD40mAb conjugate acts as an adjuvant at least in part via a direct effect on B cells , we were interested in assessing the rapidity of the antibody response induced by CD40mAb in comparison with other adjuvants. We used the model antigen, ovalbumin, in order to compare the kinetics of the induced antibody response between CD40 conjugate, MPL™ and the widely used alum adjuvant.
Female C57Bl/6 mice aged 6-8 weeks were obtained from Harlan UK Ltd and housed in accordance with strict Home Office guidelines. Ovalbumin (Sigma) was conjugated to the CD40mAb 10C8  as previously described . Four groups of 15 mice were immunised intraperitoneally with either 10 μg of conjugate, 10 μg of ovalbumin either alone, with MPL™ (10 μg, Sigma) or adsorbed onto Aluminium hydroxide (200 μg, 5). Five of the 15 mice were bled every 3 days in rotation, and anti-ovalbumin IgG endpoint titers determined by ELISA assay as previously described 
ELISA results are shown in Fig 1. CD40mAb conjugate induced an IgG response against OVA by day 7 post immunization, whereas no IgG response to OVA+MPL™ or OVA+alum was seen until day 8, and in the case of alum this was weak. The results shown are representative of a total of three experiments. Total immunoglobulin responses (including IgM) tended to arise a day earlier, but showed the same difference in kinetics between CD40 and the other two adjuvants.
CD40mAb-OVA conjugate induces a more rapid IgG response in mice than either the established adjuvant, alum, or the newer adjuvant, MPL™. How much faster the response to a CD40mAb vaccine versus an MPL adjuvanted vaccine would be in humans would need to be determined empirically. How important a more rapid response would be would depend upon the titers required to protect against a particular pathogen, as well as the window of opportunity available to prevent disease. We propose that CD40mAb conjugates may have utility in post-exposure prophylaxis when a rapid antibody response is desirable.
monophosphoryl lipid A
Toll like receptor
Enzyme linked immunosorbent assay
Ertl HC: Novel vaccines to human rabies. PLoS Negl Trop Dis. 2009, 3: e515-10.1371/journal.pntd.0000515.
Inglesby TV, O'Toole T, Henderson DA, Bartlett JG, Ascher MS, Eitzen E, Friedlander AM, Gerberding J, Hauer J, Hughes J, McDade J, Osterholm MT, Parker G, Perl TM, Russell PK, Tonat K: Anthrax as a biological weapon, 2002: updated recommendations for management. JAMA. 2002, 287: 2236-2252. 10.1001/jama.287.17.2236.
Watson B, Seward J, Yang A, Witte P, Lutz J, Chan C, Orlin S, Levenson R: Postexposure effectiveness of varicella vaccine. Pediatrics. 2000, 105: 84-88. 10.1542/peds.105.1.84.
Salzman MB, Garcia C: Postexposure varicella vaccination in siblings of children with active varicella. Pediatr Infect Dis J. 1998, 17: 256-257. 10.1097/00006454-199803000-00020.
Kohl I, Nemecek V, Summerová M, Chlíbek R, Nad'ová K, Mináriková O: Long-term protective effect of post-exposure Havrix administration during viral hepatitis Type A outbreaks. Eur J Epidemiol. 2006, 21: 893-899. 10.1007/s10654-006-9081-2.
Tagliabue A, Rappuoli R: Vaccine adjuvants: the dream becomes real. Hum Vaccin. 2008, 4: 347-349.
Paavonen J, Naud P, Salmeron J, Wheeler CM, Chow SN, Apter D, Kitchener H, Castellsague X, Teixeira JC, Skinner SR, Hedrick J, Jaisamrarn U, Limson G, Garland S, Szarewski A, Romanowski B, Aoki FY, Schwarz TF, Poppe WA, Bosch FX, Jenkins D, Hardt K, Zahaf T, Descamps D, Struyf F, Lehtinen M, Dubin G, Greenacre M: Efficacy of human papillomavirus (HPV)-16/18 AS04-adjuvanted vaccine against cervical infection and precancer caused by oncogenic HPV types (PATRICIA): final analysis of a double-blind, randomised study in young women. Lancet. 2009, 374: 301-314. 10.1016/S0140-6736(09)61248-4.
Barr TA, Carlring J, Heath AW: Co-stimulatory agonists as immunological adjuvants. Vaccine. 2006, 24: 3399-3407. 10.1016/j.vaccine.2006.02.022.
De Gregorio E, D'Oro U, Wack A: Immunology of TLR-independent vaccine adjuvants. Curr Opin Immunol. 2009, 21: 339-345. 10.1016/j.coi.2009.05.003.
Mesa C, de Leon J, Fernandez LE: Very small size proteoliposomes derived from Neisseria meningitidis: An effective adjuvant for generation of CTL responses to peptide and protein antigens. Vaccine. 2006, 24: 2692-2699. 10.1016/j.vaccine.2005.08.111.
Singh M, Chakrapani A, O'Hagan D: Nanoparticles and microparticles as vaccine-delivery systems. Expert Rev Vaccines. 2007, 6: 797-808. 10.1586/14760518.104.22.1687.
Hatzifoti C, Bacon A, Marriott H, Laing P, Heath AW: Liposomal co-entrapment of CD40mAb induces enhanced IgG responses against bacterial polysaccharide and protein. PLoS ONE. 2008, 3: e2368-10.1371/journal.pone.0002368.
Ahonen CL, Doxsee CL, McGurran SM, Riter TR, Wade WF, Barth RJ, Vasilakos JP, Noelle RJ, Kedl RM: Combined TLR and CD40 triggering induces potent CD8+ T cell expansion with variable dependence on type I IFN. J Exp Med. 2004, 199: 775-784. 10.1084/jem.20031591.
Heath AW, Chang R, Harada N, Santos-Argumedo L, Gordon J, Hannum C, Campbell D, Shanafelt AB, Clark EA, Torres R, et al: Antibodies to murine CD40 stimulate normal B lymphocytes but inhibit proliferation of B lymphoma cells. Cell Immunol. 1993, 152: 468-480. 10.1006/cimm.1993.1305.
Dullforce P, Sutton DC, Heath AW: Enhancement of T cell-independent immune responses in vivo by CD40 antibodies. Nat Med. 1998, 4: 88-91. 10.1038/nm0198-088.
Barr TA, McCormick AL, Carlring J, Heath AW: A potent adjuvant effect of CD40 antibody attached to antigen. Immunology. 2003, 109: 87-92. 10.1046/j.1365-2567.2003.01634.x.
Hatzifoti C, Heath AW: CD40-mediated enhancement of immune responses against three forms of influenza vaccine. Immunology. 2007, 122: 98-106. 10.1111/j.1365-2567.2007.02617.x.
Carlring J, Barr TA, McCormick AL, Heath AW: CD40 antibody as an adjuvant induces enhanced T cell responses. Vaccine. 2004, 22: 3323-3328. 10.1016/j.vaccine.2004.02.043.
Tang Y, Akbulut H, Maynard J, Petersen L, Fang X, Zhang WW, Xia X, Koziol J, Linton PJ, Deisseroth A: Vector prime/protein boost vaccine that overcomes defects acquired during aging and cancer. J Immunol. 2006, 177: 5697-5707.
Vonderheide RH: Prospect of targeting the CD40 pathway for cancer therapy. Clin Cancer Res. 2007, 13: 1083-1088. 10.1158/1078-0432.CCR-06-1893.
Barr TA, Carlring J, Heath AW: CD40 antibody as a potent immunological adjuvant: CD40 antibody provides the CD40 signal to B cells, but does not substitute for T cell help in responses to TD antigens. Vaccine. 2005, 23: 3477-3482. 10.1016/j.vaccine.2004.12.025.
Barr TA, Heath AW: Functional activity of CD40 antibodies correlates to the position of binding relative to CD154. Immunology. 2001, 102: 39-43. 10.1046/j.1365-2567.2001.01148.x.
AH is a Director of Adjuvantix Ltd and also holds some stock in Adjuvantix. Adjuvantix Ltd have an interest in CD40mAb based immunological adjuvants.
VB, ED, NK and LR performed the experimental work. Experiments were designed by JC, ED and AH. AH, ED and JC wrote the manuscript.
All authors have read and approved the final manuscript.
Vijay NS Bhagawati-Prasad, Evy De Leenheer contributed equally to this work.
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Bhagawati-Prasad, V.N., De Leenheer, E., Keefe, N.P. et al. CD40mAb adjuvant induces a rapid antibody response that may be beneficial in post-exposure prophylaxis. J Immune Based Ther Vaccines 8, 1 (2010). https://doi.org/10.1186/1476-8518-8-1
- Passive Immunization
- Active Vaccination
- Surface Antigen CD40