RESUMEN
In the present study, differences in hepatitis B surface antigen (HBsAg)-specific memory B-cell responses between low and high responders to hepatitis B vaccine (HepB), based on levels of antibodies against HBsAg (anti-HBs), were determined. In addition, HBsAg specific T-cell responses between high (anti-HBs level >20,000 IU/L) and low (anti-HBs level <1500 IU/L) responders were compared. Numbers of HBsAg-specific B-cells, plasma immunoglobulin G (Ig) levels, and T-cell cytokine concentrations were measured in low and high responders directly before and one month after the second booster vaccination. In advance, an Enzyme-linked Immunosorbent Spot (ELISpot) Assay was optimized for the determination of HBsAg-specific B-cell responses. The number of HBsAg-specific B-cells was significantly higher (p<0.01) in the high responder group compared to the low responder group after a booster vaccination with HepB. In addition, the plasma IgG levels and numbers of HBsAg-specific B-cells were significantly correlated (RS=0.66, p<0.01). The HBsAg-specific Th1 cell response showed the same values in the low and high responder group and did not change by the booster vaccination with HepB. However, a significant correlation (RS=0.6975, p=0.007) between the IL-13 levels and the plasma IgG levels post-booster was found. Subsequently, the IL-13 level in the high-responder group post-booster was significantly higher compared to the low-responder group. Since activation of the B-cell response after vaccination is induced by Th2 cells and IL-13 is produced by these cells, we conclude that the difference in HBsAg-specific Th2 cells is involved in determining the differences in anti-HBs level and memory B-cell numbers between low and high responders.
Asunto(s)
Linfocitos B/inmunología , Vacunas contra Hepatitis B/inmunología , Hepatitis B/prevención & control , Memoria Inmunológica , Interleucina-13/inmunología , Adolescente , Citocinas/inmunología , Ensayo de Immunospot Ligado a Enzimas , Anticuerpos contra la Hepatitis B/sangre , Antígenos de Superficie de la Hepatitis B/inmunología , Vacunas contra Hepatitis B/uso terapéutico , Humanos , Inmunización Secundaria , Inmunoglobulina G/sangre , Células Th2/inmunología , Adulto JovenRESUMEN
The progress of the Global Polio Eradication Initiative is monitored by acute flaccid paralysis (AFP) surveillance supplemented with environmental surveillance in selected areas. To assess the sensitivity of environmental surveillance, stools from (re)vaccinated elderly persons with a low seroprevalence and from wastewater were concurrently collected and analyzed in the Netherlands over a prolonged period of time. A total number of 228 healthy individuals with different levels of immunity were challenged with monovalent oral polio vaccine serotype 1 or 3. Poliovirus concentrations were determined by the titration of fecal suspensions on poliovirus-sensitive L20B cells and of sewage concentrates by L20B monolayer plaque assay. Almost half of the individuals (45%) shed poliovirus on day 3 after challenge, which peaked (57%) on day 8 with an average poliovirus excretion of 1.3 × 10(5) TCID(50) per g of feces and gradually decreased to less than 5% on day 42. The virus concentrations in sewage peaked on days 6 to 8 at approximately 100 PFU per liter, remained high until day 14, and subsequently decreased to less than 10 PFU per liter on day 29. The estimated poliovirus concentration in sewage approximated the measured initial virus excretion in feces, within 1 log(10) variation, resulting in a sensitivity of detection of 100 infected but mostly asymptomatic individuals in tens of thousands of individuals. An additional second peak observed in sewage may indicate secondary transmission missed by enterovirus or AFP surveillance in patients. This enables the detection of circulating poliovirus by environmental surveillance, supporting its feasibility as an early warning system.
Asunto(s)
Monitoreo del Ambiente/métodos , Parálisis/prevención & control , Poliomielitis/prevención & control , Poliovirus/inmunología , Vigilancia de la Población/métodos , Anciano , Anciano de 80 o más Años , Animales , Línea Celular , Monitoreo Epidemiológico , Heces/virología , Femenino , Humanos , Masculino , Ratones , Persona de Mediana Edad , Países Bajos/epidemiología , Parálisis/epidemiología , Parálisis/inmunología , Parálisis/virología , Poliomielitis/epidemiología , Poliomielitis/inmunología , Poliomielitis/virología , Poliovirus/aislamiento & purificación , Vacuna Antipolio Oral/administración & dosificación , Vacuna Antipolio Oral/inmunología , Estudios Seroepidemiológicos , Aguas del Alcantarillado/virologíaRESUMEN
Vaccination against infectious diseases ideally should provide lifelong immunity, but in many cases waning of antibody titers has been observed over time. In this study we describe the identification of antigen-specific memory B-cells in peripheral blood of persons born between 1940 and 2004 in The Netherlands. Polyclonal stimulation of either PBMCs or purified B-cells induced proliferation and differentiation of B-cells of the memory phenotype (CD19(+)/CD27(+)) into antibody secreting cells (ASC). Memory B-cells against components of bacterial vaccines (Bordetella pertussis and tetanus) as well as viral vaccines (measles and influenza) were thus identified, even in persons with low serum antibody titers. Enrichment of B-cells increased the sensitivity of memory B-cell detection when compared to PBMCs. Low, but significant correlations between numbers of antigen-specific memory B-cells and the corresponding circulating antibody titers were found for the pertussis proteins and measles virus, but not for tetanus. The identification of the numbers and specificities of peripheral memory B-cells and their relationship with circulating antibodies may be very useful to determine the long-term efficacy of vaccination.
Asunto(s)
Linfocitos B/inmunología , Vacuna contra Difteria, Tétanos y Tos Ferina/inmunología , Memoria Inmunológica , Vacuna Antisarampión/inmunología , Vacunación , Adulto , Anciano , Anticuerpos Antibacterianos/sangre , Anticuerpos Antivirales/sangre , Diferenciación Celular , Proliferación Celular , Niño , Estudios de Cohortes , Humanos , Inmunoglobulina G/sangre , Leucocitos Mononucleares/inmunología , Activación de Linfocitos , Persona de Mediana Edad , Sensibilidad y EspecificidadRESUMEN
The pathogenesis of poliovirus infection, responsible for the induction of a poliovirus-specific mucosal immune response following intraperitoneal (i.p.) inoculation of virus in mice transgenic for the poliovirus receptor (PVRTg mice), was studied. Following inoculation of poliovirus, replication was determined by increase in virus titre (TCID(50)) and by PCR of poliovirus-specific negative-strand RNA in peritoneal macrophages, mesenteric lymph nodes, Peyer's patches, duodenum, brain, kidney and liver. The presence of poliovirus antigens in several cell types was detected by immunolabelling. It was demonstrated that poliovirus replicated in the peritoneal macrophages of PVRTg mice, since the virus titre in peritoneal cells was increased compared to the titre in the inoculum. Negative-strand RNA was detected in these cells and most of the poliovirus-immunostained cells had the morphology of macrophages and expressed the macrophage-specific markers CD86 and M1/70 on their surface. Furthermore, in peritoneal lavage, poliovirus was also present in CD19(+) B cells, but not in dendritic or T cells. Moreover, poliovirus was detected in macrophage-like cells in the lamina propria of the intestine, but not in epithelial cells. Replication of poliovirus in mesenteric lymph nodes, Peyer's patches and brain was followed by excretion of virus in the faeces. This suggests that the virus is transported due to migration of macrophages from the peritoneal cavity to mesenteric lymph nodes and the lamina propria of Peyer's patches. It is likely that this route is responsible for the induction of virus-specific IgA in the gut.
Asunto(s)
Macrófagos Peritoneales/virología , Proteínas de la Membrana , Poliomielitis/virología , Poliovirus/patogenicidad , Receptores Virales/genética , Replicación Viral , Animales , Modelos Animales de Enfermedad , Humanos , Ratones , Ratones Transgénicos , Especificidad de Órganos , Poliovirus/aislamiento & purificación , Poliovirus/fisiología , Reacción en Cadena de la Polimerasa , ARN Viral/aislamiento & purificación , Receptores Virales/metabolismoRESUMEN
In view of the planned eradication of poliovirus, the suitability of transgenic mice bearing the human receptor for poliovirus (PVRtg mice) as a nonprimate animal model to study mucosal immunity against poliovirus was investigated. After intraperitoneal (ip) priming followed by ip or oral booster with live poliovirus, PVRtg mice had detectable IgA and IgG responses. The IgA response was restricted to PVRtg mice and could not be induced by oral immunization. After ip priming, PVRtg mice did shed virus in the stool, whereas control mice did not. Moreover, the amount of virus shed in the stools of PVRtg mice that had an IgA response after immunization was significantly lower than that of nonimmunized mice. A virus-specific mucosal IgA response is dependent on expression of the poliovirus receptor and is influenced by the route of immunization and the virus strain. PVRtg mice are a suitable model for the study of poliovirus-specific immunity and protection against poliovirus infection.
Asunto(s)
Anticuerpos Antivirales/biosíntesis , Inmunoglobulina A Secretora/biosíntesis , Proteínas de la Membrana , Vacuna Antipolio de Virus Inactivados/inmunología , Poliovirus/inmunología , Receptores Virales/fisiología , Animales , Femenino , Humanos , Inmunidad Mucosa , Inmunización , Inmunoglobulina G/biosíntesis , Masculino , Ratones , Ratones Endogámicos ICR , Ratones Transgénicos , Vacuna Antipolio de Virus Inactivados/administración & dosificaciónRESUMEN
In the present study, the effect of treatment with granulocyte colony-stimulating factor (G-CSF) on cellular composition of the bone marrow and the number of circulating leucocytes of granulocytopenic mice, whether or not infected with Staphylococcus aureus, was assessed. With two monoclonal antibodies, six morphologically distinct cell populations in the bone marrow could be characterised and quantitated by two-dimensional flow cytometry. Granulocytopenia was induced by cyclophosphamide or sublethal irradiation. Cyclophosphamide predominantly affected the later stages of dividing cells in the bone marrow resulting in a decrease in number of granulocytic cells, monocytic cells, lymphoid cells and myeloid blasts. G-CSF administration to cyclophosphamide-treated mice increased the number of early blasts, myeloid blasts and granulocytic cells in the bone marrow, which indicates that this growth factor stimulates the proliferation of these cells in the bone marrow. During infection in cyclophosphamide-treated mice the number of myeloid blasts increased. However, when an infection was induced in cyclophosphamide and G-CSF-treated mice, the proliferation of bone-marrow cells was not changed compared to that in noninfected similarly treated mice. Sublethal irradiation affected all bone-marrow cell populations, including the early blasts. G-CSF-treatment of irradiated mice increased only the number of myeloid blasts slightly, whereas an infection in irradiated mice, whether or not treated with G-CSF, did not affect the number of bone-marrow cells. Together, these studies demonstrated that irradiation affects the early blasts and myeloid blasts in the bone marrow more severely than treatment with cyclophosphamide. Irradiation probably depletes the bone marrow from G-CSF-responsive cells, while cyclophosphamide spared G-CSF responsive cells, thus enabling the enhanced G-CSF-mediated recovery after cyclophosphamide treatment. Only in these mice, bone marrow recovery is followed by a strong mobilisation of mature granulocytes and their band forms from the bone marrow into the circulation during a bacterial infection.
Asunto(s)
Agranulocitosis/terapia , Células de la Médula Ósea/patología , Factor Estimulante de Colonias de Granulocitos/uso terapéutico , Infecciones Oportunistas/patología , Agranulocitosis/inducido químicamente , Agranulocitosis/etiología , Animales , Antineoplásicos Alquilantes/toxicidad , Recuento de Células , Técnicas de Cultivo de Célula , División Celular/efectos de los fármacos , Ciclofosfamida/toxicidad , Femenino , Ratones , Ratones Endogámicos C57BL , Traumatismos Experimentales por Radiación/patología , Infecciones Estafilocócicas/patología , Irradiación Corporal Total/efectos adversosRESUMEN
The inactivated poliovirus vaccine (IPV) is used for protection against poliomyelitis in The Netherlands. It is not clear, however, whether IPV vaccination can lead to priming of the mucosal immune system and the induction of IgA. It has been demonstrated that IPV vaccination is able to induce strong memory IgA responses in the serum of persons who have been naturally exposed to wild-type poliovirus. This has led to the hypothesis that IPV vaccination is able to induce poliovirus-specific IgA at mucosal sites in persons who have been previously primed with live poliovirus at mucosal sites. To test this hypothesis, the kinetics of the IgA response in serum and saliva after IPV vaccination were examined in persons previously vaccinated with oral poliovirus vaccine (OPV) or IPV. ELISA and enzyme-linked immunospot assays were used for the detection of poliovirus-specific IgA responses. In addition, B cell populations were separated on the basis of the expression of mucosal (alpha4beta7 integrin) and peripheral homing receptors (L-selectin). Parenteral IPV vaccination was able to boost systemic and mucosal IgA responses in previously OPV-vaccinated persons only. None of the previously vaccinated IPV recipients responded with the production of IgA in saliva. In agreement with this finding, a large percentage of the poliovirus-specific IgA-producing lymphocytes detected in previous OPV recipients expressed the alpha4beta7 integrin. It is concluded that IPV vaccination alone is insufficient to induce a mucosal IgA response against poliovirus. In mucosally (OPV-) primed individuals, however, booster vaccination with IPV leads to a strong mucosal IgA response.
Asunto(s)
Poliomielitis/inmunología , Vacuna Antipolio de Virus Inactivados/inmunología , Poliovirus/inmunología , Adulto , Anticuerpos Antivirales/análisis , Anticuerpos Antivirales/sangre , Especificidad de Anticuerpos , Células Productoras de Anticuerpos/metabolismo , Sitios de Unión de Anticuerpos , Heces/química , Humanos , Inmunidad Mucosa/inmunología , Inmunización Secundaria , Inmunoglobulina A/biosíntesis , Inmunoglobulina A/sangre , Inmunoglobulina G/biosíntesis , Inmunoglobulina G/sangre , Inmunoglobulina M/sangre , Saliva/inmunología , Componente Secretorio/sangre , Vacunas de Productos Inactivados/inmunologíaRESUMEN
This study concerns the effect of recombinant murine granulocyte-macrophage colony-stimulating factor (GM-CSF) on the number of circulating leucocytes, activation of peritoneal macrophages and proliferation of Listeria monocytogenes in various organs of naive and leucocytopenic mice. Mice were rendered leucocytopenic by sublethal total body irradiation or cyclophosphamide treatment. GM-CSF treatment enhanced the number of granulocytes and monocytes in peripheral blood during L. monocytogenes infection in naive mice, but not in irradiated or cyclophosphamide-treated mice. In naive mice, irradiated and cyclophosphamide-treated mice, GM-CSF did not affect the course of L. monocytogenes infection in thigh muscle, spleen and liver. However, GM-CSF treatment significantly increased the number of macrophages in the peritoneal cavity of naive mice during infection; these macrophages were more enlarged and showed a higher frequency of binucleated and multinucleated cells relative to non-GM-CSF-treated mice. Together, these results demonstrated that GM-CSF increased the number of circulating granulocytes and monocytes, and the number of peritoneal macrophages during infection with L. monocytogenes in naive mice, but did not affect the course of the infection in thigh muscle, spleen or liver of these mice. In leucocytopenic mice, however, GM-CSF did not affect the number of circulating phagocytes, which explains that this factor had no effect on the proliferation of the bacteria in the various organs.
Asunto(s)
Factor Estimulante de Colonias de Granulocitos y Macrófagos/inmunología , Leucopenia/inmunología , Listeriosis/inmunología , Animales , Ciclofosfamida , Femenino , Recuento de Leucocitos , Leucopenia/etiología , Listeriosis/microbiología , Hígado/microbiología , Macrófagos Peritoneales/inmunología , Macrófagos Peritoneales/patología , Ratones , Ratones Endogámicos CBA , Músculo Esquelético/microbiología , Proteínas Recombinantes , Bazo/microbiología , Irradiación Corporal TotalRESUMEN
This study assesses the effect of granulocyte colony-stimulating factor (G-CSF) on the outcome of infection with Listeria monocytogenes or Staphylococcus aureus in mice during leukocytopenia induced by sublethal total body irradiation or cyclophosphamide treatment. The role of granulocytes during infection was assessed in the thigh muscle, spleen, and liver. G-CSF treatment in naive mice increased the number of blood granulocytes; upon infection, these numbers increased further, but G-CSF did not affect the outgrowth of bacteria in the tissues. Cyclophosphamide treatment and sublethal irradiation decreased the number of blood granulocytes, which was not affected by G-CSF treatment. However, during infection in cyclophosphamide-treated mice, G-CSF increased the number of granulocytes in the circulation and at the site of infection and decreased the number of bacteria in the tissues. Treatment with G-CSF did not affect the number of granulocytes or the course of infection in irradiated mice.
Asunto(s)
Agranulocitosis/complicaciones , Infecciones por Bacterias Grampositivas/complicaciones , Infecciones por Bacterias Grampositivas/tratamiento farmacológico , Factor Estimulante de Colonias de Granulocitos/uso terapéutico , Agranulocitosis/inducido químicamente , Animales , Ciclofosfamida/farmacología , Femenino , Rayos gamma/efectos adversos , Granulocitos/efectos de los fármacos , Granulocitos/efectos de la radiación , Listeriosis/complicaciones , Hígado/microbiología , Ratones , Ratones Endogámicos CBA , Músculos/microbiología , Proteínas Recombinantes/uso terapéutico , Organismos Libres de Patógenos Específicos , Bazo/microbiología , Infecciones Estafilocócicas/complicacionesRESUMEN
This study concerns the effect of anti-tumor necrosis factor (TNF) antibodies on the course of a sterile inflammatory reaction in the peritoneum and a generalized infection with gram-positive bacteria. Mice received an intravenous injection of rabbit anti-TNF serum or normal rabbit serum 24 h before an intraperitoneal injection of heat-killed Listeria monocytogenes or an intramuscular injection of live L. monocytogenes. The course of the leukocytes in blood and the peritoneal cavity was followed for 72 h; the infection was evaluated for 144 h. The results lead to the conclusion that anti-TNF inhibits the migration of granulocytes and monocytes from bone marrow to the circulation and from the circulation to the peritoneal cavity during an acute inflammation. Furthermore, treatment of mice with anti-TNF serum enhanced the growth of Listeria monocytogenes in thigh muscle, liver, and spleen. The results of this study indicate that treatment with anti-TNF antibodies can inhibit the development of a cellular inflammatory exudate and can have a deleterious effect on the course of an infection with gram-positive bacteria.