pneumococcal vaccine
A Streptococcus pneumoniae and vaccine technology, applied in the direction of antibacterial drugs, immunoglobulins, bacterial antigen components, etc., can solve the problem of Streptococcus pneumoniae vaccines waiting to be developed
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Embodiment 1
[0049] Embodiment 1 opsonophagocytosis bactericidal test
[0050] The opsonophagocytosis bactericidal test refers to the published standard experimental method, and the main steps are as follows: dilute the heat-inactivated serum sample and mix it with the bacteria, shake and incubate at room temperature for 30 minutes, then add the induced differentiation neutrophils and complement, The mixture was shaken and incubated at 37°C for 45 minutes, then placed on ice for 10 minutes to terminate the reaction, and then the mixture was serially diluted and plated. After overnight incubation at 37°C, the phagocytosis ratio was calculated by counting the surviving bacteria.
[0051] In the opsonophagocytosis bactericidal test, compared with the pre-immune serum, the use of rabbit serum immunized with Pbp2b, SP148 and ScpB proteins can significantly enhance the killing ability of neutrophils against Streptococcus pneumoniae. The serum dilution ratio is more than 36 times (see figure 1 ,...
Embodiment 2
[0052] The immune antigenicity detection of embodiment 2 protein
[0053] Animal experiments Adult female Balb / c mice were used. The purified Pbp2b protein, SP148 protein and ScpB protein were diluted with PBS and added with 30% volume of aluminum hydroxide adjuvant, each mouse in the experimental group was injected with 100 μL (containing 5 μg protein), and boosted immunization after 14 days and 28 days respectively . The mice in the control group were only injected with PBS and adjuvant without the above immune proteins. On the 7th day after the three immunizations, the venous blood samples of the mice were collected, and the serum was collected as the primary antibody for enzyme-linked immunosorbent assay, and the purified protein was used as the substrate to calculate the antibody titer in the serum.
[0054] The results showed that after subcutaneous injection of Pbp2b protein, SP148 protein and ScpB protein to mice (with aluminum hydroxide as adjuvant), high-titer anti...
Embodiment 3
[0055] Embodiment 3 Active immune protection test-respiratory tract infection
[0056] On the 21st day after the mice of Example 2 were immunized three times, the mice were anesthetized, and the mice were instilled with 5×10 6 40 μL of PBS suspension of Streptococcus pneumoniae. After 7 days, the mice were sacrificed, the nasal cavity was lavaged with sterile PBS and the lung tissue was taken out for grinding, the lavage liquid and the grinding liquid were diluted and coated on a flat plate, and the number and density of bacteria were calculated after overnight culture, and the experimental group was compared with the control group ( PBS suspension without Streptococcus pneumoniae) was dropped for statistical analysis.
[0057] The results showed that Pbp2b protein, SP148 protein and ScpB protein showed significant immune protection effect. Among them, in the respiratory tract infection model, the immunization of Pbp2b protein and ScpB protein significantly reduced the bacte...
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