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Recombinant Adeno-Associated Virus-Mediated Expression of Methamphetamine Antibody Attenuates Methamphetamine-Induced Hyperactivity in Mice

View Article: PubMed Central - PubMed

ABSTRACT

Methamphetamine (Meth) is one of the most frequently abused drugs worldwide. Recent studies have indicated that antibodies with high affinity for Meth reduce its pharmacological effects. The purpose of this study was to develop a technique for virus-based passive immunization against Meth effects. We generated a recombinant adeno-associated virus serotype-8 vector (AAV-MethAb) carrying the gene for a Meth-specific monoclonal antibody (MethAb). Infection of 293 cells with AAV-MethAb resulted in the expression and secretion of antibodies which bind to Meth. The viral vector was then examined in adult ICR mice. Systemic administration of AAV-MethAb resulted in long-term expression of MethAb in the serum for up to 29 weeks. Serum collected from the animals receiving AAV-MethAb retained a high specificity for (+)-Meth. Animals were challenged with Meth five weeks after viral injection. Meth levels in the brain and serum were reduced while Meth-induced locomotor activity was significantly attenuated. In conclusion, AAV-MethAb administration effectively depletes Meth from brain and serum while reducing the behavioral response to Meth, and thus is a potential therapeutic approach for Meth abuse.

No MeSH data available.


Lymphocyte counts and plasma cytokine levels in AAV-MethAb injected mice vs. those in AAV-mCherry injected mice.Mice were treated with AAV-MethAb (2.5 × 1010 VGC/animal; n = 8, i.p.) or AAV-mCherry (2.5 × 1010 VGC/animal; n = 6, i.p.). At five weeks post-infection, peripheral blood was collected for the measurement of lymphocyte counts and plasma cytokine levels. (a) Three main types of immune cells, including CD4+ T lymphocytes, CD8+ T lymphocytes, and CD19+ B lymphocytes, were counted by flow cytometry analysis. Data were expressed as mean percentage of total cell counts (lymphocytes + monocytes) ± SEM. (b) The plasma levels of six representative cytokines, including IFN- γ, TNF-α, IL1-β, IL2, IL6, and IL10, were measured by Multiplex Immunoassay using a flow cytometry-based platform (Luminex 200). Data were expressed as mean fluorescence intensity (MFI) ± SEM. No significant differences for each measured lymphocytes or cytokines were found between AAV-MethAb and AAV-mCherry injected groups.
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f4: Lymphocyte counts and plasma cytokine levels in AAV-MethAb injected mice vs. those in AAV-mCherry injected mice.Mice were treated with AAV-MethAb (2.5 × 1010 VGC/animal; n = 8, i.p.) or AAV-mCherry (2.5 × 1010 VGC/animal; n = 6, i.p.). At five weeks post-infection, peripheral blood was collected for the measurement of lymphocyte counts and plasma cytokine levels. (a) Three main types of immune cells, including CD4+ T lymphocytes, CD8+ T lymphocytes, and CD19+ B lymphocytes, were counted by flow cytometry analysis. Data were expressed as mean percentage of total cell counts (lymphocytes + monocytes) ± SEM. (b) The plasma levels of six representative cytokines, including IFN- γ, TNF-α, IL1-β, IL2, IL6, and IL10, were measured by Multiplex Immunoassay using a flow cytometry-based platform (Luminex 200). Data were expressed as mean fluorescence intensity (MFI) ± SEM. No significant differences for each measured lymphocytes or cytokines were found between AAV-MethAb and AAV-mCherry injected groups.

Mentions: To investigate whether the immune response following AAV infection can contribute to the difference in Meth-induced behavioral changes between mice infected with AAV-MethAb (2.5 × 1010 VGC/animal; i.p., n = 8) and AAV-mCherry (2.5 × 1010 VGC/animal; i.p., n = 6), we collected the whole blood at five weeks post-infection and subjected it to the measurement of lymphocyte counts and cytokine levels. There were no significant differences between these two groups of mice in the cell counts of three major lymphocyte subtypes, including CD4+ T lymphocytes (39.5 ± 2.9% vs. 37.6 ± 3.3%), CD8+ T lymphocytes (12.7 ± 1.5% vs. 11.0 ± 0.4%), and CD19+ B lymphocytes (24.6 ± 1.4% vs. 27.3 ± 3.2%) (Fig. 4a). The plasma levels of six representative cytokines, including four pro-inflammation cytokines (IFN-γ, TNF-α, IL1-β, IL6), one anti-inflammation cytokine (IL10), and one lymphocyte proliferation cytokine (IL2), were not significantly different between groups (Fig. 4b). When the cytokine levels were transformed from mean fluorescence intensity (MFI) into concentration (pg/ml), the levels of all cytokines were less than the lowest detected concentration (1 pg/ml) of the standard curve. These results suggest that AAV-MethAb infection did not alter any of the indices of immune status tested, compared to AAV-mCherry infection, nor induce systemic inflammation responses.


Recombinant Adeno-Associated Virus-Mediated Expression of Methamphetamine Antibody Attenuates Methamphetamine-Induced Hyperactivity in Mice
Lymphocyte counts and plasma cytokine levels in AAV-MethAb injected mice vs. those in AAV-mCherry injected mice.Mice were treated with AAV-MethAb (2.5 × 1010 VGC/animal; n = 8, i.p.) or AAV-mCherry (2.5 × 1010 VGC/animal; n = 6, i.p.). At five weeks post-infection, peripheral blood was collected for the measurement of lymphocyte counts and plasma cytokine levels. (a) Three main types of immune cells, including CD4+ T lymphocytes, CD8+ T lymphocytes, and CD19+ B lymphocytes, were counted by flow cytometry analysis. Data were expressed as mean percentage of total cell counts (lymphocytes + monocytes) ± SEM. (b) The plasma levels of six representative cytokines, including IFN- γ, TNF-α, IL1-β, IL2, IL6, and IL10, were measured by Multiplex Immunoassay using a flow cytometry-based platform (Luminex 200). Data were expressed as mean fluorescence intensity (MFI) ± SEM. No significant differences for each measured lymphocytes or cytokines were found between AAV-MethAb and AAV-mCherry injected groups.
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f4: Lymphocyte counts and plasma cytokine levels in AAV-MethAb injected mice vs. those in AAV-mCherry injected mice.Mice were treated with AAV-MethAb (2.5 × 1010 VGC/animal; n = 8, i.p.) or AAV-mCherry (2.5 × 1010 VGC/animal; n = 6, i.p.). At five weeks post-infection, peripheral blood was collected for the measurement of lymphocyte counts and plasma cytokine levels. (a) Three main types of immune cells, including CD4+ T lymphocytes, CD8+ T lymphocytes, and CD19+ B lymphocytes, were counted by flow cytometry analysis. Data were expressed as mean percentage of total cell counts (lymphocytes + monocytes) ± SEM. (b) The plasma levels of six representative cytokines, including IFN- γ, TNF-α, IL1-β, IL2, IL6, and IL10, were measured by Multiplex Immunoassay using a flow cytometry-based platform (Luminex 200). Data were expressed as mean fluorescence intensity (MFI) ± SEM. No significant differences for each measured lymphocytes or cytokines were found between AAV-MethAb and AAV-mCherry injected groups.
Mentions: To investigate whether the immune response following AAV infection can contribute to the difference in Meth-induced behavioral changes between mice infected with AAV-MethAb (2.5 × 1010 VGC/animal; i.p., n = 8) and AAV-mCherry (2.5 × 1010 VGC/animal; i.p., n = 6), we collected the whole blood at five weeks post-infection and subjected it to the measurement of lymphocyte counts and cytokine levels. There were no significant differences between these two groups of mice in the cell counts of three major lymphocyte subtypes, including CD4+ T lymphocytes (39.5 ± 2.9% vs. 37.6 ± 3.3%), CD8+ T lymphocytes (12.7 ± 1.5% vs. 11.0 ± 0.4%), and CD19+ B lymphocytes (24.6 ± 1.4% vs. 27.3 ± 3.2%) (Fig. 4a). The plasma levels of six representative cytokines, including four pro-inflammation cytokines (IFN-γ, TNF-α, IL1-β, IL6), one anti-inflammation cytokine (IL10), and one lymphocyte proliferation cytokine (IL2), were not significantly different between groups (Fig. 4b). When the cytokine levels were transformed from mean fluorescence intensity (MFI) into concentration (pg/ml), the levels of all cytokines were less than the lowest detected concentration (1 pg/ml) of the standard curve. These results suggest that AAV-MethAb infection did not alter any of the indices of immune status tested, compared to AAV-mCherry infection, nor induce systemic inflammation responses.

View Article: PubMed Central - PubMed

ABSTRACT

Methamphetamine (Meth) is one of the most frequently abused drugs worldwide. Recent studies have indicated that antibodies with high affinity for Meth reduce its pharmacological effects. The purpose of this study was to develop a technique for virus-based passive immunization against Meth effects. We generated a recombinant adeno-associated virus serotype-8 vector (AAV-MethAb) carrying the gene for a Meth-specific monoclonal antibody (MethAb). Infection of 293 cells with AAV-MethAb resulted in the expression and secretion of antibodies which bind to Meth. The viral vector was then examined in adult ICR mice. Systemic administration of AAV-MethAb resulted in long-term expression of MethAb in the serum for up to 29 weeks. Serum collected from the animals receiving AAV-MethAb retained a high specificity for (+)-Meth. Animals were challenged with Meth five weeks after viral injection. Meth levels in the brain and serum were reduced while Meth-induced locomotor activity was significantly attenuated. In conclusion, AAV-MethAb administration effectively depletes Meth from brain and serum while reducing the behavioral response to Meth, and thus is a potential therapeutic approach for Meth abuse.

No MeSH data available.