Compositions and Methods for Modulating Immune Response
a technology of immune response and composition, applied in the field of compositions and methods for modulating immune response, to achieve the effect of modulating immune respons
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example 1
Preparation of a H. pylori Cell Lysate
[0038]A H. pylori cell lysate was prepared following the method as described in Shin et al (supra). Briefly, H. pyloris obtained from the American Type Culture Collection (ATCC 43504) was cultured in the brucella broth (Difco laboratory, Detroit, Mich., US) including 5% fetal calf serum and antibiotics including 2.5 μg / ml amphotericin B, 10 μg / ml vancomycin, 5 μg / ml trimethoprin, and 2.5 IU / ml polymyxin B (American Sigma Chemistry Company) at 37° C., 10% CO2 with shaking at the speed of 200 rpm. Then, the culture was centrifuged at 12,000×g for 10 min, sonicated, and centrifuged. The supernatant was collected to obtain a H. pylori cell lysate. The protein concentration of the lysate was measured with the bicinchoninic acid method (Pierce, Rockford, Ill.).
example 2
Immunization of Hens and Preparation of Yolk Lyophilized Powders
[0039]The above-described H. Pylori lysate obtained in Example 1 (protein concentration, 200 μg / ml) was mixed with the same volume of a complete Freund adjuvant (Difco laboratory, Detroit, Mich., US). The mixture was intramuscularly injected to hens (25 week age, Brown Leghorn hens, n=15). Each hen had 4 positions at the leg muscle to accept the injection; the dosage for each position is 250 μl. After the first injection, a mixture of the H. Pylori lysate and the same volume of an incomplete Freund adjuvant was intramuscularly injected to the hens every other week for three times to boost the immune response. After one month following the last boost, the eggs laid by the hens were collected and the yolk was removed and lyophilized to obtain yolk lyophilized powders, called “IgY-HP.” IgY-HP was confirmed to have an adequate title of antibodies for quality control.
example 3
Animal Experiments
[0040]3.1 Animals
[0041]Female BALB / c mice, 6-week old, were used in the following experiments. They were purchased from BioLASCO Taiwan Co., Ltd. (Taipei, Taiwan), and maintained in accordance with the procedures approved by the Food Industry Development Research institute (Hsinchu, Taipei). Briefly, mice after purchased were observed for two weeks and those with abnormal behaviors (fearing light, dehydration, etc.) or having weights out of the average weight±2 standard deviation were discarded. Each of the cages was placed with a layer of sawdust (the American NEPCO Corporation, Warrensburg, N.Y.), which was sterilized before placement and replaced twice times per week. The feed was PMI 5001 specific for experimental mice purchased from Purina (USA). The mice were kept at 23±2° C., 50±10% relative humidity, and 12:12 (hours) light dark cycle.
[0042]3.2 Feeding
[0043]The mice were randomly divided into two groups, the IgY-HP experimental group and the water control g...
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