猪流行性腹泻病毒免疫的最新知识

Latest Knowledge on PEDv Immunity, PED猪流行性腹泻病毒免疫的最新知识Michael P. Murtaugh迈克尔·莫特夫 Albert Rovira Bob Morrison Cheryl Dvorak Suzanne Stone Diem Ngo Sally Robinson Mike Rahe Xiong Wang Dane Goede Iowa Select Farms Ian Levis Shamus Brown Prestage Farms Matthew Turner Sara Hough Schafer Farms Tim Snider

National Pork Board国家猪肉局

从左到右：苏珊·斯通，谢丽尔·德沃夏克，卡西·郭，迈克·拉赫，安·霍伊布克，莎莉·罗宾逊，本杰明·威尔，迪姆·傲，玛丽娜·菲格雷多

Left to right: Suzanne Stone, Cheryl Dvorak, Cassie Guo, Mike Rahe, Ann Hoybook, Sally Robinson, Benjamin Wier, Diem Ngo, Marina Figueiredo

Conclusions结论 PEDV protection requires mucosal immune response.猪流行性腹泻病毒保护需要黏膜免疫反应。 Mucosal immunity requires oral administration and is based on IgA antibodies in the intestinal lumen.粘膜免疫需要口服给药，并基于在肠腔的IgA抗体。 One feedback induces immunity and viral clearance with or without clinical signs.反饲产生免疫力和病毒清除，可能有或没有临床症状。

Conclusions结论 Feedback induces seroconversion, but nucleocapsid antibody titers decline over time.反饲引起血清转换，但随着时间的推移，核蛋白壳抗体滴度下降。 Milk antibodies are IgA against S1 and S2 spike protein fragments. We predict that these are protective.奶中抗体IgA抗S1和S2刺突蛋白片段。我们预测，这些是有保护性的。 Duration of PEDV immunity is≥4 months.猪流行性腹泻病毒免疫的时间是≥4个月。

Immunity in the Porcine Gut

猪肠道免疫力

淋巴集结

远距离肠道粘膜

胸管激活淋巴滤泡周边血液其他外分泌组织激活淋巴滤泡

Oral Administration口服

0

7

14

21

25

ImmunizeDennis Foss,博士论文 PhD Thesis, UMN免疫

Days

Examine检测

Experimental Model

实验模型

Readout– ASC读数-ASC

Oral Exposure Primarily Induces Mucosal IgA

口服感染主要是诱导粘膜IgA

ASC/106总细胞

Dennis Foss, PhD Thesis, UMN

固有层

脾脏

Oral Exposure is Required for Mucosal Response

粘膜反应需要通过口腔感染Oral Antigen: Mucosal IgA口腔抗原：粘膜IgA

SQ抗原：系统IgG SQ Antigen: Systemic IgG

抗CT-B ASC 106总细胞

对照Dennis Foss, PhD Thesis, UMN

处理/给药途径

Oral Induction of Anti-CT-B IgA and IgG ASC口腔诱导的抗CT-B IgA和IgG ASC3000 Anti-CT-B ASC/106 cells 2500 2000 1500 1000 500 0 control 0 CT-B 7 CT-B+ CT 14 21 Anti-CT-B ASC/106 cells

Lamina Propria固有层 IgA IgG

10

Spleen脾IgA IgG

7.5

5.0

2.5

0

control

CT-B

CT-B+ CT

25

Immunize免疫

Days

Examine检查

Experimental Model实验模型

Readout– ASC读数-ASCDennis Foss. Michael Murtaugh University of Minnesota

PEDV Immunity Assumptions猪流行性腹泻病毒免疫假设 Oral administrati

on in sows induces durable intestinal immunity.母猪口服诱导持久肠道免疫力。 Lactogenic immunity can be passed to suckling piglets to protect them against disease.猪乳中产生的的免疫力可以传递给哺乳仔猪以保护他们免受疾病。 Effective vaccines are not available.有效的疫苗市场上没有。 Immunity by feedback is a useful prevention tool.反饲产生的免疫一个非常有效的预防手段。

Porcine Epidemic Diarrhea Virus猪流行性腹泻病毒 Coronavirus family冠状病毒科 Enveloped,+

sense, ssRNA genome, appx 28,000 nt in length.包被+感应，单链RNA基因组，约28,000 NT长。 Closely related to TGEV与猪传染性胃肠炎密切相关 Spike (S1 and S2 fragment) and M are present on the surface.刺突 (S1和S2片段)和M是存在于表面上。

S1/S2

N MFigure from D. Song and B. Park. Porcine epidemic diarrhoea virus: a comprehensive review of molecular epidemiology, diagnosis, and vaccines. Virus Genes (2012) 44:167-175.

ELISA Antigens酶联免疫吸附检测抗原KDa 199 N S1 S2

His-tagged PEDV N, S1, and S2 proteins were expressed and purified有组氨酸标记的 PEDV N,S1和S2蛋白被表达和纯化

122

86

40 31

样本数量50 100 150 200 250 300 350 400 0

Number of Samples

0.2

Look at ELISA Values

酶联免疫反应读值0.5

ELISA S/P ELISA S/P值

POS

NEG阴性

阳性

0-0.1 0.1-0.2 0.2-0.3 0.3-0.4 0.4-0.5 0.5-0.6 0.6-0.7 0.7-0.8 0.8-0.9 0.9-1.0 1.0-1.1 1.1-1.2 1.2-1.3 1.3-1.4 1.4-1.5 1.5-1.6 1.6-1.7 1.7-1.8 1.8-1.9 1.9-2.0

Sow Shedding母猪散毒周数

WEEKS WEEK 1 WEEKS WEEK 1 1+3 1+3

1. Non-clinical pigs shed PEDV无临床表现的猪散毒 1. All sows immune at week 3所有的母猪在第3周免疫 1. Massive exposures unnecessary不需要大量感染 WEEKS 1+3 WEEK 1 WEEKS 1+3 WEEK 1

Ct值

一周

四周

Short-lived antibody presence in sows and piglets母猪和仔猪体内短期存在的抗体

ELISA S/P比值

周数

Antibodies to nucleocapsid are present at week 4, but diminished at week 9抗核衣壳抗体存在于第4周，但在第9周减弱。

猪乳汁中的猪流行性腹泻病毒抗体是IgAIgG IgA

PEDV Antibodies in Milk are IgA

OD值

3 Sow Herds: Milk contains IgA猪乳中含有IgA

Antibodies are against S1 and S2抗体抗S1和S2 These antibodies are important for protection这些抗体起重要保护作用