the present invention is related to the use of the hepatitis b virus surface antigen as an immunoenhancer in nasal immunizations , with vaccine formulations resulting from the combination of this antigen and other vaccine antigens that benefit from this property and the application of this property of the hepatitis b virus surface antigen and the use of formulations in the field of vaccines . it is also related with multivalent formulations , specifically for nasal administration resulting from the application of this property of the hbsag , favoring an increase in the immune response of other antigens present in the formulations . vaccine formulations for the nasal administration of this invention together with the hepatitis b virus surface antigen may contain one or more protein antigens of a soluble nature , receiving an immunoenhancing effect due to its co - administration with hbsag . these may be : tetanus toxoid , diphtheria toxoid or protein - polysaccharide conjugates where the saccharide part corresponds to a vaccine candidate anti haemophilus influenzae type b , the c polysaccharide of neisseria meningitidis , the vaccine polysaccharide of pneumococcus pneumoniae , or in general , one or more soluble proteins of vaccines of interest either purified or obtained recombinantly . also within the aim of this invention are the multivalent formulations for nasal administration in which the hepatitis b virus surface antigen is combined with a vaccine candidate from inactivated microorganisms which receives an immunoenhancing effect because of its co - administration with hbsag . the vaccine antigen may be bordetella pertussis whole - cell , which receives an immunoenhancing effect because of its co - administration with hbsag or other vaccine antigens of the same nature alone , or integrating complex combinations of antigens . other vaccine antigens that may be contained are the present inactivated or attenuated vaccine candidates . vaccine formulations for nasal administrations related with the present invention may contain an n number of antigens from microorganisms of different species ranging from n = 1 to n = 20 , of antigenic nature among those previously described , with a final volume and antigen amounts for inoculation ranging from 50 microliters to 2 milliliters , and from 0 . 1 micrograms to 2 mg , respectively , depending on the size and the species to be immunized . formulations of the present invention may be solubilized in pbs , saline solution , water for injection or in any buffer solution used in medical practice that allows for antigen stability , in an antigen concentration lying within the possible combinations of mass and volume previously described . likewise , the antigenic components can be mixed with hbsag according to the candidates of interest in relation to vaccination age or to multivalent candidates based on any other premise , where one , two or more antigen types described above are presented either lyophilized or administered in drops , sprays or pulverization . vaccine formulations of the present invention can be used to attain an effective immunization in humans or animals as a preventive or therapeutic treatment . fig1 . kinetics of igg response of an experiment where inoculations were achieved on days 0 , 14 , 28 y 87 and bleedings on days — 10 , 21 , 35 , 42 , 84 and 97 . ( a ) kinetics of serum igg response against hbsag ; ( b ) kinetics of serum igg response against tt ; ( c ) kinetics of serum igg response against td and ( d ) kinetics of serum igg response against bp . tables 1a , b , c and d : results of the statistical analyses of the comparisons between groups of the corresponding figures . fig2 . vaginal iga response on day 97 . ( a ) iga response against hbsag ; ( b ) iga response against tt ; ( c ) iga response against dt and ( d ) iga response against whole - cell bp . tables 3a , b , c and d : statistical analyses . fig3 . lung iga response on day 97 . ( a ) iga response against hbsag ; ( b ) iga response against tt ; ( c ) iga response against dt and ( d ) iga response against whole - cell bp . tables 3a , b , c and d : statistical analyses . fig4 . evaluation of serum igg response against individual proteins of bp , after the administration of tetravalent formulations by the intra - peritoneal or nasal routes . optical density levels generated by individual sera of each group immunized with nasal or intra - peritoneal tetravalent formulations ( groups 7 and 13 , respectively ), after ( a ) a third inoculation and ( b ) a fourth inoculation . fig5 . evaluation of the proliferative activity against antigens found in the nasal tetravalent formulation of the example 1 , administered individually and within the tetravalent formulation . fig6 . enhancing activity of hbsag on neisseria meningitidis ompc . the combination of both antigens significantly increased the anti - ompc response . with the aim of evaluating the antibody response generated after the nasal administration of several formulations containing different types of antigens together with or without hbsag , an experiment was designed with 126 female balb / c mice , 8 to 10 weeks of age , divided into 13 groups : groups 1 to 11 with 10 animals each , and groups 12 and 13 with 8 animals each . all mice were immunized at days 0 , 14 , 28 and 87 and bled at days — 10 , 21 , 35 , 42 , 84 and 97 . the dose of each antigen administered per mouse is shown below : group route dose of antigen per group group 1 : ( nasal ) 1 5 μg of hbsag + 10 μg of dt group 2 : ( nasal ) 5 μg of hbsag + 3 . 2 uop of bp * group 3 : ( nasal ) 5 μg of hbsag + 10 μg of tt group 4 : ( nasal ) 5 μg of hbsag + 10 μg of dt + 3 . 2 uop of bp * group 5 : ( nasal ) 5 μg of hbsag + 10 μg of dt + 10 μg of tt group 6 : ( nasal ) 5 μg of hbsag + 3 . 2 uop of bp * + 10 μg of tt group 7 : ( nasal ) 5 μg of hbsag + 10 μg of dt + 10 μg of tt + 3 . 2 uop of bp * group 8 : ( nasal ) 5 μg of hbsag group 9 : ( nasal ) 10 μg of dt group 10 : ( nasal ) 3 . 2 uop of bp * group 11 : ( nasal ) 10 μg of tt group 12 : ( ip ) 2 5 μg of hbsag + 0 . 125 mg of al ( oh ) 3 group 13 ψ : ( ip ) 5 μg of hbsag + 49 . 26 μg of dt + 29 . 07 μg of tt + 8 . 0 uop of bp * + 0 . 125 mg of al ( oh 3 ) ( nasal ) 1 group with nasal immunization ( ip ) 2 group with intra - peritoneal immunization * uop : units of opacity ; in each case equal amounts of uop of both bp strains described above , were used . ψ it can be observed that group 13 had the same hbsag dose as the nasal groups . however , the doses of the remaining antigens here were higher because a commercial vaccine formulation was used as the control . this vaccine contained specific doses of each antigen that could not be changed . these correspond to the micrograms presented in the table . hence the amount of tt , dt and bp used through the intranasal route are 3 , 5 and 2 . 5 fold less respectively . # the same amounts of hbsag were nonetheless used . determinations of igg antibody response against the hepatitis b virus surface antigen indicated that a week after the third administration , in the group nasally immunized with hbsag , dt , tt and bp ( group 7 ) it was significantly higher than group 8 , which only received hbsag in pbs by the same route . this behavior was repeated on day 42 ( fig1 a ). the other nasally immunized groups ( groups 1 - 6 ) showed the same behavior as group 7 , developing igg levels that are significantly greater than those obtained with hbsag in pbs after three doses , except for group 2 , which had a higher , but not significant ( p & gt ; 0 . 05 ) response on day 35 . nevertheless , this group did have a very significant difference on day 42 , which can be explained by a broad dispersion of the titers on day 35 . the groups nasally immunized with formulations containing hbsag and bp ; hbsag , bp and dt ; and hbsag , bp and tt ( groups 2 , 4 y 6 respectively ) had significantly higher igg responses than those shown by group 8 after the second dose . it is good to point out that on day 35 , the anti - hbsag igg response of the group that was nasally immunized with the hb - dtp mixture was not significantly different from that of the homologous group immunized through an intra - peritoneal injection ( group 13 ). in the same way , after four doses , statistically similar values were obtained between nasal immunized groups and the ip immunized group using the antigen combination ( fig1 ). anti - hbsag titers generated in the group immunized with the heberbiovac hb ® vaccine ( group 12 ) were significantly higher than those obtained with any nasal group . this result is due to a higher immunogenicity of hbsag when it is administered by the intra - peritoneal ( ip ) route with respect to intramuscular and subcutaneous routes . this characteristic of the ip route has been shown in our experiments . in spite of this characteristic of the anti - hbsag response , the inoculation of 250 μl of the vaccine per mouse , corresponding to the dose of 5 μg of hbsag — equivalent to nasal dose — makes it necessary to use of ip route . nevertheless , it has been observed in older animals that the response generated by the nasal inoculation equals that induced by intra - peritoneal injection . an example of this was seen in the second experiment ( see example 2 ) though the nasal candidates were different . despite a lower titer intensity generated by the nasal route , it has been recently observed that anti - igg2a response levels induced in group 2 were significantly higher than those shown by group 12 ( data not shown ). this demonstrates a modulating effect of bp on the anti - hbsag antibody response , introducing a qualitative change in the response , favoring the th1 - like response , which is evidenced by the igg subclass profile . this response is characterized by a higher production of igg2a antibodies with respect to the common profile generated by the vaccine adjuvanted in alum . as observed in fig1 , the appearance and sustainment of the nasal and intra - peritoneal anti - hbsag responses were very similar , with a similar increase and decrease in time . a stronger immunoenhancing effect of hbsag , dt and bp on tetanus toxoid was evidenced after their nasal administration in the group 7mixture . after the second , third and fourth inoculations , the anti - tt igg levels of this group showed a highly significant increase with respect to group 11 , immunized with tetanus toxoid in pbs by the same route . the other groups nasally vaccinated with mixtures containing tt , specifically groups 3 , 5 and 6 , showed higher antibody levels to those induced in group 11 after each bleeding . in the case of group 3 , an antibody level approximately 100 times higher than group 11 was attained after the second dose . this is highly relevant because this is the first demonstration of the immunoenhancing activity of hbsag on a soluble antigen such as tt . as observed in the anti - hbsag response , tetanus toxoid also increased the anti - hbsag response , showing that we are in the presence of a synergistic interaction with respect to a cross - enhancement of immunogenicity . this type of phenomenon had already been observed for hbsag and hbcag and other viral nucleocapside antigens , but never with a soluble antigen . tt is not immunogenic by the nasal route , which is evidenced in group 11 , immunized with the toxoid in pbs . hence , this effect was not expected with hbsag and tt . anti - tt response was strengthened significantly more when hbsag , tt and bp were formulated together ( group 6 ) producing anti - tt responses 1000 times stronger than those generated by intranasal immunization with tt in pbs ( group 11 ). this additional increase was not obtained by the mixture of dt , hbsag and tt ( group 5 ). this formulation did not produce a significant change , but showed a statistical behavior similar to that of the combination of tt and hbsag in respect to anti - tt response ( fig1 b ). titers of groups 6 and 7 were not significantly different . both these groups having in common the presence of bp and hbsag , induced responses that were 1000 times higher . therefore , we can say that both the surface antigen and bp produce an immunoenhancement of the anti - tt response that increases titers to very significant levels in the first case and to highly significant levels with the mixture of both . since all groups of intranasal combinations in the experiment were immunized with hbsag , it is important to define whether the enhancing effect of bp is independent of hbsag or whether both have a synergistic enhancing activity . it can be affirmed with respect to tt , that the number of antigens present in all formulations studied did not affect its immunogenicity , but it was increased still more , reaching highly significant levels ( fig1 b ). the tt - specific igg levels induced in groups 7 and 13 , both immunized with the 4 antigens by the intranasal and intra - peritoneal administration respectively , did not show significant differences . responses were , however , obtained through the intranasal route for certain groups that were higher , although not significantly so , compared to those obtained in group 13 ( fig1 b ). although the groups immunized with hbsag and tt ( group 3 ) and with hbsag , dt and tt ( group 5 ) did not have different igg titers to those obtained in group 13 , a week after the second dose , they had a significantly lower response after the third and fourth doses . this result evidenced that it is important to add bp to the mixture in order to increase anti - tt response to levels similar to those obtained after ip administration . nevertheless , it should also be taken into account that amounts of tt 3 times higher were administered by the ip than by the nasal route . the nasal administration of dt together with hbsag , tt and bp ( group 7 ) led to a considerable increase in dt immunogenicity , compared to the response reached by group 9 , in which dt was administered in pbs ( fig1 c ). the same behavior was shown in the rest of the groups nasally immunized with dt combinations ( groups 1 , 4 and 5 ) in highly significant increases in anti - dt igg levels . only group 1 did not show a significant superior result after second dose . however , 35 and 42 days after the third dose , the response of group 1 was highly significant compared to the group immunized with diphtheria toxoid in pbs ( fig1 c ). the comparison of anti - dt igg levels between nasally immunized groups with formulations containing dt and the group with the intra - peritoneal administration , the tetravalent commercial vaccine ( group 13 ) showed an anti - tt response - like behavior . in other words , the response was enhanced 100 fold in groups containing hbsag but not bp , whereas the addition of bp allowed to increase the responses still more , even to levels higher than those generated by the ip route with the dpt - hb vaccine . this behavior of dt reproduces the effect found for the other soluble protein , tt . hence , we can affirm that hbsag has a strong immunoenhancing effect on both soluble proteins and vice versa . in the case of dt , a synergistic activity was also evidenced with regard to the cross - enhancement of the immune response to the antigens into the mixtures ; hence , as it was explained before , anti - surface antigen igg response was enhanced by the addition of dt . once more the need of adding bp enabling to increase the anti - dt response one - hundred times , to levels similar to those obtained after ip administration , was demonstrated . it must be recalled that the response is similar to that obtained through the ip route although in that case the amount of dt was five times larger . [ heading - 0089 ] anti - b pertussis whole cell igg antibody response in the serum all anti - bp responses were characterized by their strength and quick increase to levels near saturation ( fig1 d ). in spite of the resulting homogeneity , statistical analysis revealed the possibility of the generation of significant differences between groups . after a third administration the group nasally immunized with the tetravalent combination generated a significantly higher response than that obtained in the group immunized with bp alone through the same route ( group 10 ). however , this effect disappeared after the fourth inoculation since it rapidly reached a state of titer saturation . the other groups nasally immunized with formulations containing bp together with one or more antigens had strong responses , that were significantly to highly significant greater in the case of the group given hbsag and bp on day 35 , evidencing that the immunoenhancing effect of hbsag is also produced in inactivated cells , in addition to the known effect on soluble proteins . both groups 2 and 4 maintained this statistical difference on day 42 , while groups 6 and 7 ( having administered the more complex combinations ) did not differ from group 10 . a comparison of all intranasal groups with the ip group showed no significant differences after the second and third doses , evidencing that high titers can be obtained by nasal immunization with the whole cell . [ heading - 0094 ] determination of mucosal response of the nasal multivalent formulations of hbsag . taking into account that a stronger response to the nasally administered antigens depends on whether a strong response can be generated at the mucosal and systemic levels , we determined iga antibody response in vaginal and lung ravages in the immunized groups that are described in example 1 . after the fourth administration , on day 100 , hbsag - specific iga response induced in vaginal lavages of the nasally immunized group with the tetravalent formulation , was not significantly different from that found in mice exclusively immunized with hbsag through the same route . it is important to point out that at this time , serum anti - hbsag igg titers were not different either because of the strong response generated in the group immunized with hbsag in pbs by boosting . however , a 20 % higher seroconversion was found in the first group . although no differences were found , it should be highlighed that such a large amount of antigens does not affect vaginal anti - hbsag response . on the other hand , when comparing iga levels induced in the groups receiving the combination of the four antigens either through the intranasal or intra - peritoneal routes ( groups 7 and 13 respectively ) it was demonstrated that the levels of predominant antibodies in vaginal secretions , developed in the group immunized through the mucosal route were significantly higher than those of the parenterally immunized group . it should be stressed that the intra - peritoneal inoculation with hbsag formulations results in a vaginal iga level , that although low , its value is higher than those obtained by the sc and im routes ( data not shown ). that is why the response found in vaginal lavages after ip immunization were significant . in general , intranasal immunization exceeded ip , as expected . the determination of vaginal tt - specific iga levels after four administrations evidenced that the group nasally immunized with the combination of hbsag , dt , bp and tt ( group 7 ) generated a greater and highly significant response compared to those induced in the group only immunized with tt , one of which had 0 % seroconversion . the other groups nasally inoculated with tt in different antigenic mixtures had the same behavior . from this result , and bearing in mind the response of group 3 ( containing hbsag and tt ), it is possible to state that hbsag enhanced vaginal anti - tt iga response at the same level as the other mixtures ( fig2 b ). all groups inoculated by the intranasal route showed greater and highly significant responses compared to the group immunized with hb - dtp through intra - peritoneal injection . this group only had a 14 % seroconversion . thus , it can be affirmed that not all antigens administered through the ip route give a vaginal response . it seems to be an exclusive property of certain antigens , including hbsag . anti - dt iga response induced in the group immunized with the nasal tetravalent formulation as well as in the groups immunized with other nasal combinations containing dt were greater ( p & lt ; 0 . 001 ) than that generated in group 9 which was given a control preparation of dt in pbs ( fig2 c ). moreover , the response generated by the group given the four antigens nasally was also greater than that generated in the group of the intra - peritoneal - administered tetravalent vaccine ; a similar behavior to that of tt . in general , it was seen that the mucosally immunized groups developed vaginal iga levels that were significantly higher than those of the group given the four antigens ip . anti - dt iga levels belonging to group 1 did not differ from those obtained by the groups having higher immunogenicity , evidencing , as formerly confirmed for tt , that hbsag induced a highly significant enhancement of the vaginal anti dt response compared to group 9 , which was administered the toxoid in the pbs ( fig2 c ). it is possible , as we commented before , that this result may not have a direct benefit on the protection against tetanus and diphtheria . however , after evaluating the tt and dt models it can be suggested that the nasally administered combination of hbsag with antigens of similar nature — not particulate but soluble — from other pathogens , free or anchored to these proteins , could generate an increase in the immune response at the vaginal level . vaginal anti - bp response was divided into two levels , a lower one in which the group immunized with bp in pbs was located , and another upper one corresponding to the immunized groups with all studied combinations . a greater and highly significant anti - bp response was obtained by combined immunization ( all containing hbsag ). nevertheless , it should be noted that the response generated by bp alone induced levels of nearly 1 : 100 , unlike tetanus and diphtheria toxoid which scarcely seroconverted and only reached strong responses after their immunization in the antigen mixtures . in the case of bp the ip inoculation was not observed to generate any vaginal iga responses . therefore , it had a tt - like behavior , in which one generated by the ip route , a negligible iga response in vaginal ravages . the other two antigens , hbsag and dt alone generated weak but significant responses having 70 and 60 % of seroconversion respectively . in the same way that vaginal responses are important in protecting against one of the pathogens , a response at the respiratory tract is very important for pathogens bp and corynebacterium diphtheriae . this does not exclude studies on anti - tt and anti - hbsag responses for general knowledge about the level of mucosal immunologic activation after intranasal immunization with the tetravalent formulation of group 7 . after determining the hbsag - specific iga response in lung ravages , it was concluded there were no significant differences between that generated by the group immunized nasally with the tetravalent combination ( group 7 ) and that generated by the group which only received hbsag by the same route ( group 8 ). it is important to point out that during the evaluation of iga levels in the lungs , there were similar serum igg levels in groups immunized either with the mixture or with hbsag in pbs . the strong boosting effect due to the fourth administration could trigger anti - hbsag iga levels in group 8 . however the response was strong and not affected with that generated by hbsag in pbs by the administration of a large amount of antigens ( dt , tt and bp ) evidencing the high capacity of the route ( fig3 a ). in the same figure it can be observed that the ip route , does not induce a significant response in lung lavages and the comparison with the intranasal groups widely favours the latter ( p & lt ; 0 . 001 ). the anti - tt iga response of the tetravalent nasal formulation was much higher ( p & lt ; 0 . 001 ) than that obtained with tt in pbs by the same route , or with intra - peritoneal administration together with hbsag , dt , and bp , in alum . the responses shown by these two latter control groups were almost negligible ( fig3 b ). the combination of diphtheria toxoid with nasally administered hbsag , tt and bp , enormously enhanced its immunogenicity . this is based on the fact that results of the determinations of anti - dt iga antibody responses indicated that there were highly significant differences ( p & lt ; 0 . 001 ) between the group immunized with the combination of the four antigens by the mucosal route ( group 7 ) and groups 9 and 13 , corresponding to dt in pbs nasally administered and the control of the tetravalent vaccine through the intra - peritoneal route , respectively . in the case of group 9 , no mouse was positive to dt - specific iga in a dilution of 1 : 100 . in the ip group only one positive mouse was detected . hence , this poor response strongly contrasts with that obtained by the tetravalent mixture , while there is also a contrast in the response obtained against tt in the homologous groups ( fig3 c and 3b ). after comparing anti - bp iga responses developed in the group immunized with the tetravalent combination by the intranasal route ( group 7 ) and the group exclusively immunized with inactivated bp cells by the same route ( group 10 ), no differences were found between them . however , when each response from these groups was compared to that generated by the group given the alum - adjuvanted mixture of four elements through the intra - peritoneal route , the superiority of both iga responses was highly significant , demonstrating once again that only a mucosal inoculation favors strong increases in the iga response detected in lung ravages ( fig3 d ). although the anti - bp response in lung lavages was not enhanced with the mixture as in vaginal lavages , the increase in antigen number was not found to negatively affect immunogenicity . it should be noted that the anti - bp response was very high in groups 7 and 10 , with titers of a geometric mean of up to 10 4 . this result shows that bp is an excellent immunogen by the intranasal route inducing lung iga responses . the effect observed in antibody levels was of a maximum narrowing of intervals , evidencing their saturation . since lung lavages were only performed as of group 7 and onward , group 2 was not compared to group 10 to study the effect of combining hbsag and bp , but we assume , considering titer levels and the characteristic of the response , the differences would be very small , if any . [ heading - 0122 ] comparison of antibody response against the proteins fha and pertussis toxin after nasal and systemic administrations of the formulations of groups 7 and 13 of example 1 . because of previous reports mentioned in the specification suggest a lower ability of the intranasal route in order to elicit a response against the individual proteins of bp : fha and pertussis toxin , the evaluation of the response against them was carried out in groups 7 and 13 , corresponding to the tetravalent formulation administered by the nasal and parenteral routes , respectively . this evaluation was achieved after three and four inoculations . the statistical analysis of the response of the evaluated bleedings demonstrated there were no significant differences between the nasal groups and the parenteral ones . therefore , we could conclude that in the nasal tetravalent formulation the induction continues even after the inoculation of a 2 . 5 times lower amount of bp ( fig4 ). in order to study the spleen &# 39 ; s proliferative response generated by the antigens of example 1 , groups 7 to 13 were selected on day 100 , extracting the spleen from at least four mice per group , making a lymphocytes pool , cultured in the presence of the antigens they had been immunized with . the results of the evaluation of the proliferative capacity of the antigens nasally administered in a tetravalent formulation are individually highlighted in the fig5 . intra - peritoneal groups served as the control route . as a result of this experiment it was evidenced that it is possible to obtain significant cellular response against all antigens present in the preparation , and even higher in some of them , from the nasal administration of multiple formulations of antigens , fig5 . [ heading - 0127 ] hbsag also act as an enhancer of the immunogenicity of the protein complex from the outer membrane vesicles of neisseria meningitidis ( ompc ). in order to explore whether hbsag is able to enhance the response against co - administered antigens , an experiment was carried out in which groups of 8 balb / c mice of 8 to 10 weeks of age were immunized with hbsag , ompc , and the corresponding control groups of the antigens alone . this experiment evidenced that the surface antigen was able to significantly enhance the immune response against ompc and vice versa ( fig6 ). other co - inoculated antigens received a similar effect on their immunogenicity because of the enhancing activity of the surface antigen . some of them are inactivated virus , attenuated microorganisms and viral . surface proteins , in addition to soluble proteins and bacterins . some of the multiple combinations , which have demonstrated the immunogenicity of the individual components and which have generated an increased response against a high percentage of the antigens present within them are shown in the following table . they can be formulated as a whole in a liquid or lyophilized form for nasal administration . 1 - hb - d 2 - hb - p 3 - hb - t 4 - hb - dp 5 - hb - dt 6 - hb - pt 7 - hb - dpt 8 - hb -( hib ) 9 - hb -( ompc ) 10 - hb -( attenuated or inactivated virus , native or recombinant ) 11 - hb - dpt - hib 12 - hb -( ipv ) 13 - hb - dpt - hib -( ipv ) among the antigenic combinations that could be mixed with hbsag , antigens could be selected which could be formulated according to their application in age groups of : elderly persons , adolescents or newborn children , and according to the kind of mucosal disease . included here are the antigens that enter through the mucosa , in which mucosal immunity is important . they can be selected according to the use of the antigen mixtures to prevent sexual , respiratory or mouth - intestinal diseases , according to the risk groups or to the travelers &# 39 ; needs , according to the organ ( for instance : hbv , hcv and hav ), or according to the kind of disease ( for instance : chronic sexually - transmitted diseases , viral sexually - transmitted diseases , etc . . . )
the invention relates to a mucosal surface antigen which is used to promote and increase in the immune response against co - administered antigens in the formulations out line in the invention . said novel formulations are obtained from the dual use of the surface antigen as an immunostimulatory agent and , at the same time , as a vaccine antigen . in this way it is possible to obtain multiple formulations of the hepatitis b surface antigen and heterologous antigens , with immunogenicity levels similar to those obtained following parenteral administration and with a reduction in components that can dispense with the use of nasal adjuvants , thereby converting same antigens into elements that can promote an increase in the response to the other co - administered antigens . said novel use of the hepatitis b virus surface antigen and the resulting antigen formulations can be used in the pharmaceutical industry as therapeutic and preventive vaccine formulations .