A treatment strategy for cancer is the suppression of tumor growth by directing an immune response to the tumor vessels, which will destroy the tissue. In this thesis we describe the development of a vaccine that targets antigens expressed around angiogenic vasculature in most solid tumors. These antigens are alternative spliced extra domains of glycoproteins present in the extracellular matrix; e.g. the extra domain-B (ED-B) and extra domain-A (ED-A) of fibronectin and the C-domain of tenascin-C (TNCC). We show that it is possible to break self-tolerance and induce a strong antibody response against ED-B by vaccination. Furthermore, tumor growth was inhibited and the changes observed in the tumor tissue were consistent with an attack of the tumor vasculature by the immune system. For clinical development of therapeutic vaccines, targeting self-molecules like ED-B, a potent but non-toxic biodegradable adjuvant is required. The squalene-based Montanide ISA 720 (M720) in combination with CpG DNA fulfilled these requirements and induced an equally strong anti-self immune response as the preclinical golden standard Freund’s adjuvant. We have further characterized the immune response against ED-B generated with the adjuvant M720/GpG.  The ED-B vaccine also inhibited tumor growth in a therapeutic setting in a transgenic mouse model of pancreatic insulinoma in which tumorigenesis was already initiated. Furthermore, antibodies against ED-A and TNCC could be induced in mice and rabbits. We analyzed the expression of ED-A in breast tumors of transgenic MMTV-PyMT mice, a metastatic breast cancer model, with the aim to use this model to study the effect of an ED-A vaccine on metastasis. We also detected ED-B in canine mammary tumor tissue. Therefore vascular antigens might also represent potential therapeutic targets in dogs.  All together our preclinical data demonstrate that a vaccine targeting tumor blood vessels is a promising new approach for cancer treatment.

This thesis examined the evidence, policy decision-making, and implementation of HPV vaccination in Canada as a case study to explore normalization versus individualized decision making in public health programs. Mixed methods were used: a systematic review, content analyses and policy document analysis. Overall, the scientific evidence supported an effect of vaccination against HPV infection and precancerous cervical lesions, but evidence regarding cervical cancer incidence or mortality is lacking. Scientific and medical communities appeared optimistic about the vaccine, but cautious about its readiness for routine implementation. Policy decision-making was initially cautious, but shifted towards active program implementation, possibly related to the availability of federal funding. The educational materials and media coverage both sent clearly normalizing messages about HPV vaccination. The discussion suggests that HPV vaccination might be more suited to an individualized than population approach, but many factors coincided to promote its implementation, in Canada, within a traditional public health model.

As respiratory syncytial virus (RSV) is a respiratory pathogen that causes significant morbidity and mortality in infants, there has always been great interest in the development of a vaccine. In the 1960s, children were immunized with formalin-inactivated (FI)-RSV vaccines. Not only did these vaccines fail to prevent infection, but in most cases they resulted in enhanced disease upon subsequent exposure to the virus. In the intervening years, studies in mice have led to the hypothesis that the enhanced disease is due to an aberrant Th2-biased immune response. Thus, we hypothesized that formulating FI-RSV vaccines with a Th1 promoting adjuvant, such as CpG oligoeoxynucleotides (ODN), would result in the induction of protective immunity against RSV without risk of deleterious effects. We observed in calves that parenterally delivered FI-bovine RSV (BRSV) formulated with CpG ODN resulted in a shift towards a Th1-biased or more balanced immune response that was protective against BRSV.<p> As RSV infects the lung mucosa, vaccines that induce mucosal immunity are desirable. Parenterally delivered vaccines typically induce systemic immunity with low mucosal immune response levels, whereas mucosally delivered vaccines induce systemic and mucosal immunity. However, upon mucosal delivery there is an increased chance of vaccine components being degraded or washed away prior to the induction of immunity. Thus, we added polyphosphazenes (PP) to our mucosal vaccine formulations. PP are synthetic polymers that form non-covalent complexes with other vaccine components, increasing their stability. Intranasally delivered FI-BRSV co-formulated with CpG ODN and PP performed better than FI-BRSV alone, or FI-BRSV formulated with either adjuvant individually, in terms of inducing protective immunity against BRSV in mice. Furthermore, mice that received intranasally-delivered FI-BRSV or BRSV F protein co-formulated with CpG ODN and PP developed higher levels of immunity and protection than mice that received parenterally delivered vaccines. Because of the similarities between BRSV and HRSV, co-formulation of intranasally delivered HRSV vaccines with CpG ODN and PP could prove important in the development of a safe vaccine against HRSV in humans.

Immunization has been used as an upstream, protective measure in public health for decades. Although immunization programs have been introduced in Pakistan, new and emerging infectious disease remains a concern in the country. The province of Sindh, Pakistan is of special concern because of its large rural population.<p>The purpose of this study was to: 1) determine and compare complete and age-appropriate immunization uptake in children 12 to 36 months and birth to 9 months, respectively living in Sindh, Pakistan in 1994 and 1997; and 2) determine the correlates of complete and age-appropriate immunization in children 12 to 36 months and birth to 9 months, respectively living in Sindh, Pakistan in 1997.<p>This study reviewed data that was collected as part of the School Nutrition Program (SNP) and Family Health Project (FHP) in 1994 and 1997, respectively. Analyses included immunization data on 1877 children from the SNP survey and 1694 children from the FHP survey.<p>Females were found to have higher statistically significantly age-appropriate uptake than males in 1997 (p=0.015). Complete immunization status was also found to vary significantly by district of residence in 1994 and 1997 (p<0.001). Both complete and age-appropriate immunization status was found to decrease from 1994 to 1997.<p>Multivariable logistic regression revealed that not owning a radio, electricity, or bicycle was indicative of lower odds of complete immunization uptake (OR<1, p<0.05). Other correlates predictive of lower odds of complete immunization included owning a water pump (OR=0.360), not having a Lady Health Worker (LHW) visit the home (OR=0.489), living in a kucha house (OR=0.637), and living in Tharparkar (OR=0.290), Badin (OR=0.599), or Mirpur Khas (OR=0.271).<p>A similar regression analysis revealed childs sex, ownership of a refrigerator, and having heard of contraception to be correlates of age-appropriate immunization (p<0.05). Females had higher odds of age-appropriate immunization (OR=1.851) compared to males. Not having a refrigerator was indicative of lower odds (OR=0.079). Not having heard of at least one type of contraception was a predictor age-appropriate immunization (OR=1.925).

Immunization has been used as an upstream, protective measure in public health for decades. Although immunization programs have been introduced in Pakistan, new and emerging infectious disease remains a concern in the country. The province of Sindh, Pakistan is of special concern because of its large rural population.<p>The purpose of this study was to: 1) determine and compare complete and age-appropriate immunization uptake in children 12 to 36 months and birth to 9 months, respectively living in Sindh, Pakistan in 1994 and 1997; and 2) determine the correlates of complete and age-appropriate immunization in children 12 to 36 months and birth to 9 months, respectively living in Sindh, Pakistan in 1997.<p>This study reviewed data that was collected as part of the School Nutrition Program (SNP) and Family Health Project (FHP) in 1994 and 1997, respectively. Analyses included immunization data on 1877 children from the SNP survey and 1694 children from the FHP survey.<p>Females were found to have higher statistically significantly age-appropriate uptake than males in 1997 (p=0.015). Complete immunization status was also found to vary significantly by district of residence in 1994 and 1997 (p<0.001). Both complete and age-appropriate immunization status was found to decrease from 1994 to 1997.<p>Multivariable logistic regression revealed that not owning a radio, electricity, or bicycle was indicative of lower odds of complete immunization uptake (OR<1, p<0.05). Other correlates predictive of lower odds of complete immunization included owning a water pump (OR=0.360), not having a Lady Health Worker (LHW) visit the home (OR=0.489), living in a kucha house (OR=0.637), and living in Tharparkar (OR=0.290), Badin (OR=0.599), or Mirpur Khas (OR=0.271).<p>A similar regression analysis revealed childs sex, ownership of a refrigerator, and having heard of contraception to be correlates of age-appropriate immunization (p<0.05). Females had higher odds of age-appropriate immunization (OR=1.851) compared to males. Not having a refrigerator was indicative of lower odds (OR=0.079). Not having heard of at least one type of contraception was a predictor age-appropriate immunization (OR=1.925).

As respiratory syncytial virus (RSV) is a respiratory pathogen that causes significant morbidity and mortality in infants, there has always been great interest in the development of a vaccine. In the 1960s, children were immunized with formalin-inactivated (FI)-RSV vaccines. Not only did these vaccines fail to prevent infection, but in most cases they resulted in enhanced disease upon subsequent exposure to the virus. In the intervening years, studies in mice have led to the hypothesis that the enhanced disease is due to an aberrant Th2-biased immune response. Thus, we hypothesized that formulating FI-RSV vaccines with a Th1 promoting adjuvant, such as CpG oligoeoxynucleotides (ODN), would result in the induction of protective immunity against RSV without risk of deleterious effects. We observed in calves that parenterally delivered FI-bovine RSV (BRSV) formulated with CpG ODN resulted in a shift towards a Th1-biased or more balanced immune response that was protective against BRSV.<p> As RSV infects the lung mucosa, vaccines that induce mucosal immunity are desirable. Parenterally delivered vaccines typically induce systemic immunity with low mucosal immune response levels, whereas mucosally delivered vaccines induce systemic and mucosal immunity. However, upon mucosal delivery there is an increased chance of vaccine components being degraded or washed away prior to the induction of immunity. Thus, we added polyphosphazenes (PP) to our mucosal vaccine formulations. PP are synthetic polymers that form non-covalent complexes with other vaccine components, increasing their stability. Intranasally delivered FI-BRSV co-formulated with CpG ODN and PP performed better than FI-BRSV alone, or FI-BRSV formulated with either adjuvant individually, in terms of inducing protective immunity against BRSV in mice. Furthermore, mice that received intranasally-delivered FI-BRSV or BRSV F protein co-formulated with CpG ODN and PP developed higher levels of immunity and protection than mice that received parenterally delivered vaccines. Because of the similarities between BRSV and HRSV, co-formulation of intranasally delivered HRSV vaccines with CpG ODN and PP could prove important in the development of a safe vaccine against HRSV in humans.

Analytical methods used clinically in Sweden for detection and quantification of fetal hemoglobin (HbF) in maternal blood are either the microscopic method Kleinhauer Betkes test (KBT) or high performance liquid chromatography. A more modern alternative to detect and quantify HbF+ erythrocytes is flow cytometry. The aim of this project was therefore to evaluate the commercial kit "Fetal Cell Count kit" using flow cytometry. The kit used two antibodies; one directed against the specific γ-chain of HbF protein and the other directed against the intracellular enzyme carbanhydrase (CA), which is found in all erythrocytes in adults. The resulting data showed good precision, sensitivity and linearity. A reference interval based on male blood donors was determined to &lt;0.1 % HbF+ erythrocytes and &lt;1.3 %F-cells. The kit is well suited to detect and quantify F-cells. It could be used as a important tool to follow-up patients withβ-thalassemia and sickle cell anemia. However the kit was not as useful for detection and quantification of HbF+ erytrocytes in fetomaternal hemorrhage induced by Rhimmunization.

No description available.

Thesis (M.A.)--West Virginia University, 2001. / Title from document title page. Document formatted into pages; contains viii, 47 p. : ill. Vita. Includes abstract. Includes bibliographical references (p. 29-35).

Thesis (Ph.D.)--Virginia Commonwealth University, 2009. / Prepared for: Dept. of Human Genetics. Title from title-page of electronic thesis. Bibliography: leaves 166-195.