RESUMO

Background: CD4+ T cells, which are often referred as T-helper cells, play a central role through secreting various cytokinesto enhance immune defense to pathogen. CD8+ T cells, which are called cytotoxic T lymphocytes (CTLs), provide potentdefenses against virus infection and intracellular pathogens by killing the targets cells directly. In our previous researches,the conventional and semi-quantitative PCR were used to detect the goose CD4 and CD8α. However, the semi-quantitativeRT-PCR only detect the relative amount of gene transcription. Quantitative PCR assay was more sensitive than conventionalPCR assay, and quantitative PCR assay has a lower limit of sensitivity.Materials, Methods & Results: Contrast to conventional assays, the detection of amplicons by quantitative RT-PCR couldbe visualized as the amplifi cation progressed. This effect has provided a great deal of insight into the kinetics of the reaction and it is the foundation of kinetic of real-time qPCR. The analysis of gene transcription by qPCR has proven to bean attractive method due to its potential for increasing laboratory throughput, simultaneous processing of several samplesas well as more reliable instrumentation. With those in mind, the real-time quantitative reverse transcription PCR (qRTPCR) methods for the detection of goose CD4 and CD8α transcripts were reported here for the fi rst time. With this assay,it is possible to carry out a rapid quantitative analysis of goose CD4 and CD8α transcripts over a wide linear range, withan unknown template.CD8 is expressed on the membrane of T cells either as an αα-homodimer or αβ-heterodimer. Sinceboth forms of CD8 have α chain, the transcription levels of CD8 can be monitored by detecting CD8α mRNA expression.Assays were based on the DNA sequence of goose CD4 [GenBank: JX902315], CD8α [GenBank: KC476104], and β-actin[GenBank: M26111]. qPCR was carried out in quadruplicates in a total volume of 20 µL containing...

Assuntos

RESUMO

Background: CD4+ T cells, which are often referred as T-helper cells, play a central role through secreting various cytokinesto enhance immune defense to pathogen. CD8+ T cells, which are called cytotoxic T lymphocytes (CTLs), provide potentdefenses against virus infection and intracellular pathogens by killing the targets cells directly. In our previous researches,the conventional and semi-quantitative PCR were used to detect the goose CD4 and CD8α. However, the semi-quantitativeRT-PCR only detect the relative amount of gene transcription. Quantitative PCR assay was more sensitive than conventionalPCR assay, and quantitative PCR assay has a lower limit of sensitivity.Materials, Methods & Results: Contrast to conventional assays, the detection of amplicons by quantitative RT-PCR couldbe visualized as the amplifi cation progressed. This effect has provided a great deal of insight into the kinetics of the reaction and it is the foundation of kinetic of real-time qPCR. The analysis of gene transcription by qPCR has proven to bean attractive method due to its potential for increasing laboratory throughput, simultaneous processing of several samplesas well as more reliable instrumentation. With those in mind, the real-time quantitative reverse transcription PCR (qRTPCR) methods for the detection of goose CD4 and CD8α transcripts were reported here for the fi rst time. With this assay,it is possible to carry out a rapid quantitative analysis of goose CD4 and CD8α transcripts over a wide linear range, withan unknown template.CD8 is expressed on the membrane of T cells either as an αα-homodimer or αβ-heterodimer. Sinceboth forms of CD8 have α chain, the transcription levels of CD8 can be monitored by detecting CD8α mRNA expression.Assays were based on the DNA sequence of goose CD4 [GenBank: JX902315], CD8α [GenBank: KC476104], and β-actin[GenBank: M26111]. qPCR was carried out in quadruplicates in a total volume of 20 µL containing...(AU)

Assuntos

RESUMO

Background: Newcastle disease (ND), caused by avian paramyxovirus serotype 1 (APMV-1), also known as Newcastle disease virus (NDV), is regarded as one of the two most devastating diseases of poultry with the characteristics of serious disease and high flock mortality. It causes severe economic losses in domestic poultry, especially in chickens. At present, there were no effective drugs to treat this disease and the main method to control ND was vaccination. Moreover, new strains of virus resistant to chemotherapy continue to emerge, so does the need for a safe and effective vaccine. The immune adjuvant can make vaccine generate a strong immune response providing long-term protection against infection. With commonly usage of some immune adjuvants (e.g. mineral oil and aluminium hydroxide), many problems were occurred, such as side effects, strong local stimulation and carcinogenesis, together with complicated preparation, or failure to increase immunogenicity of weak antigen and so on. Botanical polysaccharides, as a new type of adjuvant or immunopotentiator, had become the hot research area because of their less side effects and no toxicity. The purpose of this research was to observe whether Chuanminshen violaceum polysaccharide (CVPS) possessed synergistical immunoenhancement, and offer the theoretical evidence for developing potential new-type adjuvant. Materials, Methods &a

Background: Newcastle disease (ND), caused by avian paramyxovirus serotype 1 (APMV-1), also known as Newcastle disease virus (NDV), is regarded as one of the two most devastating diseases of poultry with the characteristics of serious disease and high flock mortality. It causes severe economic losses in domestic poultry, especially in chickens. At present, there were no effective drugs to treat this disease and the main method to control ND was vaccination. Moreover, new strains of virus resistant to chemotherapy continue to emerge, so does the need for a safe and effective vaccine. The immune adjuvant can make vaccine generate a strong immune response providing long-term protection against infection. With commonly usage of some immune adjuvants (e.g. mineral oil and aluminium hydroxide), many problems were occurred, such as side effects, strong local stimulation and carcinogenesis, together with complicated preparation, or failure to increase immunogenicity of weak antigen and so on. Botanical polysaccharides, as a new type of adjuvant or immunopotentiator, had become the hot research area because of their less side effects and no toxicity. The purpose of this research was to observe whether Chuanminshen violaceum polysaccharide (CVPS) possessed synergistical immunoenhancement, and offer the theoretical evidence for developing potential new-type adjuvant. Materials, Methods &a

RESUMO

Background: Newcastle disease (ND), caused by avian paramyxovirus serotype 1 (APMV-1), also known as Newcastle disease virus (NDV), is regarded as one of the two most devastating diseases of poultry with the characteristics of serious disease and high flock mortality. It causes severe economic losses in domestic poultry, especially in chickens. At present, there were no effective drugs to treat this disease and the main method to control ND was vaccination. Moreover, new strains of virus resistant to chemotherapy continue to emerge, so does the need for a safe and effective vaccine. The immune adjuvant can make vaccine generate a strong immune response providing long-term protection against infection. With commonly usage of some immune adjuvants (e.g. mineral oil and aluminium hydroxide), many problems were occurred, such as side effects, strong local stimulation and carcinogenesis, together with complicated preparation, or failure to increase immunogenicity of weak antigen and so on. Botanical polysaccharides, as a new type of adjuvant or immunopotentiator, had become the hot research area because of their less side effects and no toxicity. The purpose of this research was to observe whether Chuanminshen violaceum polysaccharide (CVPS) possessed synergistical immunoenhancement, and offer the theoretical evidence for developing potential new-type adjuvant. Materials, Methods &a

Background: Newcastle disease (ND), caused by avian paramyxovirus serotype 1 (APMV-1), also known as Newcastle disease virus (NDV), is regarded as one of the two most devastating diseases of poultry with the characteristics of serious disease and high flock mortality. It causes severe economic losses in domestic poultry, especially in chickens. At present, there were no effective drugs to treat this disease and the main method to control ND was vaccination. Moreover, new strains of virus resistant to chemotherapy continue to emerge, so does the need for a safe and effective vaccine. The immune adjuvant can make vaccine generate a strong immune response providing long-term protection against infection. With commonly usage of some immune adjuvants (e.g. mineral oil and aluminium hydroxide), many problems were occurred, such as side effects, strong local stimulation and carcinogenesis, together with complicated preparation, or failure to increase immunogenicity of weak antigen and so on. Botanical polysaccharides, as a new type of adjuvant or immunopotentiator, had become the hot research area because of their less side effects and no toxicity. The purpose of this research was to observe whether Chuanminshen violaceum polysaccharide (CVPS) possessed synergistical immunoenhancement, and offer the theoretical evidence for developing potential new-type adjuvant. Materials, Methods &a

RESUMO

Background: Newcastle disease (ND), caused by avian paramyxovirus serotype 1 (APMV-1), also known as Newcastle disease virus (NDV), is regarded as one of the two most devastating diseases of poultry with the characteristics of serious disease and high flock mortality. It causes severe economic losses in domestic poultry, especially in chickens. At present, there were no effective drugs to treat this disease and the main method to control ND was vaccination. Moreover, new strains of virus resistant to chemotherapy continue to emerge, so does the need for a safe and effective vaccine. The immune adjuvant can make vaccine generate a strong immune response providing long-term protection against infection. With commonly usage of some immune adjuvants (e.g. mineral oil and aluminium hydroxide), many problems were occurred, such as side effects, strong local stimulation and carcinogenesis, together with complicated preparation, or failure to increase immunogenicity of weak antigen and so on. Botanical polysaccharides, as a new type of adjuvant or immunopotentiator, had become the hot research area because of their less side effects and no toxicity. The purpose of this research was to observe whether Chuanminshen violaceum polysaccharide (CVPS) possessed synergistical immunoenhancement, and offer the theoretical evidence for developing potential new-type adjuvant. Materials, Methods & Results: 200 three-yellow chickens at one day of age were randomly divided into four groups and reared in separated pens. On 7 days old, the average maternal serum hemaglutination inhibition (HI) antibody titer was less than 3 log2, all chickens of each group (the average body weight (BW) was 120 g) were vaccinated with ND live vaccine through nasal drip and eye-drop. At the same time, the chickens in three CVPS groups (high, medium and low doses of CVPS) were orally administered with 0.5 mL of CVPS at concentrations of 100 mg/kg BW, 50 mg/kg BW, 25 mg/kg BW respectively, once a day for fi ve successive day, in negative control group (NC), with 0.5 mL of physiological saline, once a day for fi ve successive day. On days 14, 21 and 28, the serum antibody titer, erythrocyte-C3b receptor rosette rate (E-C3bRR), erythrocyte-C3b immune complex rosette rate (E-ICRR), peripheral lymphocyte proliferation and peripheral CD4+/CD8+ ratio were measured. The results showed that the antibody titer, E-C3bRR, elimination rate of immune complex and peripheral lymphocyte proliferation in three CVPS groups and peripheral CD4+/CD8+ ratio in medium dosage of CVPS group were significantly higher (P < 0.05) than those in control group throughout the process of whole experiment almost. Discussion: The CVPS not only improved the E-C3bRR and accelerated the elimination rate of CIC, but also induced higher antibody titer, peripheral lymphocyte proliferation and peripheral CD4+ /CD8+ ratio in chickens vaccinated against ND live vaccine. This indicated that CVPS possessed immune-enhancement efficacy of ND live vaccine and might be expected as a candidate of new-type adjuvant.

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