ABSTRACT
The feasibility of a single-shot, low-dose vaccination against pandemic influenza was investigated. The immunogenicity and safety of whole inactivated, cell culture-derived H5N1 virus plus CoVaccine HT™ as adjuvant was tested in various animal species. In ferrets, doses of 4.0 and 7.5 µg H5N1 (NIBRG-14; A/Vietnam/1194/04; clade 1) without adjuvant gave low geometric mean haemagglutination inhibition (HI) titres (GMTs) of 21-65 three weeks after intramuscular (IM) injection. The addition of 0.25-4 mg CoVaccine HT™ resulted in GMTs of 255-1470 corresponding with 4-25-fold increases. A second immunization caused GMTs of 8914-23,525 two weeks later, which confirmed strong priming. One out of 8 ferrets injected with antigen alone and 5 out of 32 ferrets injected with adjuvanted H5N1 demonstrated minimal transient, local reactions and two animals immunized with adjuvanted H5N1 exhibited increased body temperature one day after injection. In macaques, 5 µg H5N1 with CoVaccine HT™ or aluminium hydroxide as adjuvant elicited GMTs of 172 and 11, respectively three weeks later. A second immunization resulted in GMTs of 1751 and 123, respectively four weeks later. Analysis of cross-reactivity of antibodies after the first immunization with NIBRG-14 adjuvanted plus CoVaccine HT™ revealed GMTs of 69 against NIBRG-23 (A/turkey/Turkey/1/05; clade 2.2) and 42 against IBCDC-RG-2 (A/Indonesia/5/05-like; clade 2.1.3) while titres with aluminium hydroxide were <10. After the second immunization with CoVaccine HT™, GMT against NIBRG-23 was 599 and against IBCDC-RG-2 254, while those with aluminium hydroxide were 23 and 13, respectively. No local or systemic adverse events were detected in macaques. Safety of 5 µg H5N1 plus 0, 2 or 4 mg CoVaccine HT™ was investigated in a repeated dose study in rabbits. Groups of 6 or 9 male and female animals were immunized IM three times at three week intervals. None of the animals exerted treatment-related adverse reactions during the study or at necropsy 3 or 4 days after treatment. We concluded that a low dose of whole inactivated influenza virus plus CoVaccine HT™ is a promising, single-shot vaccine against pandemic influenza.
Subject(s)
Influenza A Virus, H5N1 Subtype/immunology , Influenza Vaccines/administration & dosage , Influenza Vaccines/immunology , Vaccination/methods , Adjuvants, Immunologic/administration & dosage , Animals , Antibodies, Viral/blood , Cross Reactions , Female , Ferrets , Fever/chemically induced , Hemagglutination Inhibition Tests , Immunization, Secondary/methods , Influenza Vaccines/adverse effects , Injections, Intramuscular , Macaca , Male , Rabbits , Skin Diseases/chemically induced , Vaccines, Inactivated/administration & dosage , Vaccines, Inactivated/adverse effects , Vaccines, Inactivated/immunologyABSTRACT
The influence of blood-membrane interaction on human peripheral blood monocyte tumor necrosis factor-alpha (TNF), interleukin-6 (IL-6), and interleukin-8 (IL-8) secretion was measured during hemodialysis of end-stage renal disease patients by in vitro stimulation of whole blood with lipopolysaccharide. Monocyte TNF and IL-6 secretion in vitro was reduced 30 min after start of dialysis session. In contrast, cellular IL-8 secretion did not change during hemodialysis. Comparison of the results of three different membranes indicates that the bioincompatibility of the dialysis membrane was reflected in both leukocytopenia and reduction of cellular TNF secretion. During treatment of normal whole blood in an ex vivo dialysis closed-loop circuit, the ability of monocytes to release TNF, IL-6, and IL-8 in vitro remained constant. This indicates that the reduced IL-6 and TNF secretion during standard hemodialysis was not due to a direct effect of contact between dialysis membranes and monocytes, but rather was a result of redistribution within the patients' leukocyte pool.
