vaccination against haemonchus contortus

Vaccination against Haemonchus contortus

Dr. ISHFAQ MAQBOOLMVSc SCHOLAR

DIVISION OF PARASITOLOGY

Introduction

Haemonchus contortus

Ubiquitous and highly pathogenic.

Mostly in tropical and sub-tropical countries (Kalita et al., 1978)

Temperate regions with focal areas of similar climatic conditions (Waller et al., 2004; Waller et al., 2006)

In India - 3 species of Haemonchus - H. contortus, H. longistipes and H. similis

(Sood, 1981)

Life cycle

A female lays around 5000-10000 eggs/day

0.05 – 0.07 ml/parasite/day leading to anaemia. (Malviya et al.,1979; Soulsby, 1982)

Punctiform haemorrhages in abomasal mucosa

Hypoproteinemia, bottle jaw

↓ Digestion and absorption of proteins, calcium and phosphorus (Sood, 1981)

Decreased feed conversion, malnutrition, loss of appetite, low fertility indices and in certain cases death of young animals

(Quiroz et al., 1984)

Economic losses are incurred through morbidity, mortality and increased investment due to cost of preventative as well as curative treatments

(Miller et al., 2006)

GI parasites in sheep alone cause annual losses ranging from US $ 42 million to US $ 222 million

(Waller, 2006) Losses due to GI parasites in sheep was estimated to be US $500 million in Australia

and $946 million in New Zealand (Emery, 1991;Vlassoff and

McKenna, 1994)

Estimated treatment cost alone for H. contortus per year in India was US $103 million (McLeod, 2004)

Chronic haemonchosis is very common and of considerable economic importance (Newton and Munn, 1999)

Economic Importance

Anthelminthic resistance (Kaplan et al., 2005; Burke and Miller, 2006)

Demand for “clean and green” animal products free of residual chemicals and growth promoters (Waller, 2003)

Increasing toxicity and persistence of chemical residues in the environment (Wolstenholme et al., 2004)

Immunization with irradiated L3 stage larvae (Mulligan et al., 1961)- Failure to protect young lambs (Smith and Christie, 1979)

Need for Immunoprophylaxis

How it works?

Adequate control can be achieved with vaccine efficacies of about 80% (Barnes et al. 1995)

Natural or Surface or E/S antigens (Emery et al., 1999; Alunda et al.,

2003)

Hidden or Concealed antigens (Smith, 1999; Knox et al., 1999)

Types of Antigens

Schallig et al 1997 identified adult 15 and 24 kDa as immunogenic polypeptides in ES products of H.contortus

Gomez-Munoz et al., 1995 identified and purified 26 kDa somatic antigen but Cornelissen, 1996 concluded that 26-kDa Somatic and the 24-kDa E/S antigens of Haemonchus contortus one and the same differing in glycosylation

Karanu et al.,1993 identified Cysteine proteinases in excretory-secretory (ES) products with estimated molecular weights (Mr) of 32, 35, 38, and 40 kDa

Natural antigens

Gut antigens of fourth larval stage (L4) and adult parasites

Contortin: (Munn, 1977 ) Helical polymeric structure attached to the luminal surface of the intestinal cells

Aminopeptidase H11 glycoprotein: (Munn et al.,1992 ) It runs on (SDS-PAGE) as a doublet with a mean value of 110 kDa It has microsomal (membrane) aminopeptidase M and microsomal aminopeptidase A activities

attributable to distinct isoforms (Smith et al 1997, Graham et al 1993)

Haemonchus galactose-containing glycoprotein complex: (Smith et al., 1994 ) It contains aspartyl, metallo- and cysteine proteinases. Major component is a family of four zinc

metallo-endopeptidases, designated MEPs 1–4, MEP3 being most dominant (Smith et al., 1999)

Hidden antigens…

p52 and p46: Mixture of two gut surface proteins of 52 and 46 kDa obtained by affinity chromatography

(Sharp et al., 1992)

Cysteine proteinases and Glutamate dehydrogenase : The expression of the GDH and cysteine protease encoding genes coincides with the onset

of blood-feeding

Cysteine proteinase: Cathepsin-B family expressed in intestine of adult worms

(Rehman and Jasmer, 1999;Skuce et al., 1999) Cysteine proteases in H. contortus are capable of digesting Hb, fibrinogen, collagen and

IgG (Knox et al., 1993)

GDH: Predominant portion of S3 TSBP of 60 kDa (Knox et al. 1995)

Expressed in the cytoplasm of the intestinal cells (Skuce et al., 1999)

Hidden antigens…

Fractionate and vaccinate approach (Emery and Wagland, 1991)

Gut antigens H11 (Munn et al., 1993) H-gal-GP (Smith et al., 1994, 1999) Cysteine protease (Knox et al., 1995)

Use of immune serum 15 and 24 kDa ES Ag (Schallig et al., 1994)

Use of ASC probes HcsL3 (Meeusen et al., 1995)

Identification of Immunodominant antigens

Antigen Animal specifications

Protection Immunoglobulin Reference

r15/24 LambsAdults

55% FECR49% FECR

Low IgA,IgG,IgE level Vervelde et al.,2002

rHcp26/23 All age groups No significant protection Strong IgG and Lymphoproliferative response

Leticia et al .,2010

rH11 6 m old lambs No significant protection - Sexton and Zawadski .,2005

H-gal-GP Blackface × Leicester male lambs, aged 9 months

No significant protection High levels of immunoglobulins

Smith et al.,2003Cachet et al.,2010

Cysteine ProteinaseHc58

Sheep

Goat (7-8 m)

29-38%

Slight protection

Strong correlation

Increase in IgA,IgG and Mucosal IgA level

Redmond and Knox.,2004

Muleke et al.,2007

rHc23 Lambs (6-7 m) >80% FEC85% WB reduction

Increased level of IgA,IgG,IgE

Fawzi et al.,2015

Vaccination with recombinant Ag’s

Recombinant Antigen

Reason/s for Failure Reference

15 and 24 kDa Redundancy Newton and Meeusen.,2003

p26/23 Lack of glycosylation Leticia et al .,2010

H-gal-GP Inappropriate glycosylation and/or conformational folding

Smith et al., 2003aSmith et al., 2003b

H-gal-GP Failure to identify the protective component of the complex, or the requirement for the antigen(s) to be presented in a complex form

Newton and Meeusen.,2003

H11

Inappropriate glycosylation and/or conformational folding

Haslam et al., 1996Munn et al.,1997 Newton and Meeusen.,2003

H11 Lack of additional components that are present in native H11-enriched extract

Roberts et al., 2013

Cysteine proteinase

Inappropriate folding Redmond and Knox.,2004

Failure of vaccination with rAg

Antigen/dose Adjuvant Animal specifications

Efficacy Immunoglobulin Reference

15 and 24 kDa50-100µg

DDA Lambs 72.9%FEC reduction82.2% WB reduction

Strong correlation Schallig & van Leeuwen, 1997; Schallig et al., 1997

15 and 24 kDa50-100µg

DDA Sheep 82% WB reduction in 9 m old77% in 6m old0% in 3m old

Strong correlation Kooyman et al.,2000

p26/23 DDA Lambs >60% FEC>61.6% WB reduction

Strong correlation Domınguez-Torano et al., 2000

Cysteine protease Aluminum hydroxide

Lambs 77%FEC reduction and 47% WB reduction

Significantly higher local and systemic ES specific IgA and IgG responses.

Boisvenue et al., 1992Bakker et al., 2004De Vries et al. (2009)

Hc-sL320 µg

Aluminum hydroxide

Five month old lambs

61% FEC reduction69% WB reduction

No difference Jacobs et al., 1999

Hc23100 µg

Al(OH)3

BacterialLambs 70.67%,85.64% FEC

67.1% , 86% WB reductionrespectively

Strong correlation Fawzi et al., 2014

Vaccination with native natural Ag

Antigen Animal specifications

Efficacy Immunoglobulinlevels

Reference

Contortin Lambs 75% reduction in worm burden Strongly correlated Munn et al., 1987

p52 and p46 Lambs 78% FEC reduction

33%WB reduction

- Smith et al., 2000

p52 and p46 14-month-

old goats

34% FEC reduction and 53%

WB reduction

- Jasmer et al., 1993

Cysteine proteinaseS3 TSBPS3 TSBPS3 TSBPPurified cysteine proteaseS3 TSBPPurified cysteine protease

Sheep

Goats

Lambs

Lambs

Lambs

95% FECR,50% WBR

77% FECRand 47%WBR

89% FECRand 68%WBR

Comparable results

Significant protection

Significant protection

Predominant IgG2 response

High IgG levels

High levels of antibody

Knox et al., 1995

Knox et al.,1999

Ruiz et al., 2004

Knox et al., 2005

Redmond et al., 2006

Molina et al., 2015

Vaccination with native hidden Ag

Antigen/dose/adjuvant

Animal specifications


Reference

H11 50 µg FCA/FIA

Young Dorset lambs 78% FECR 83% WB reduction

Significant correlation

Tavernor et al. 1992

H11 fraction 0.35µg FCA/FIA

Young merino lambs 70.7% FECR70% WB reduction


Munn et al., 1993

H11 140µgH11 200µg

4 month old lambs 94.6% FEC reduction86.5% and 93.5% male and female WB reduction respectively


Smith et al., 1994

H11 from resistant strains150µg FCA/FIA

Lambs 99% FECR90% WB reduction


Newton et al., 1994

H11 (4 isolates)150µg FCA/FIA

Lambs 82–96% FECR 55.9–93.8%WB reduction


Newton et al., 1994

H11 50 µg FCA/FIA .

Pregnant ewes 91% FECR86% WB reduction

Significantly higher, Colostral transfer

Andrews et al., 1995

H11 40 µg Vax saponin

Lambs 99.9% FECR93.6% WB reduction


Roberts et al.., 2013

Vaccination with native H11

Antigen/dose/adjuvant

Animal specifications


Reference

H-gal-GP 200 µg in FCA

Suffolk x Dorset 3-5 m old lams

89% FECR 69.5% WB reduction


Smith & Smith .,1996

H-gal-GP 100µg FCA/FIA

5-6 m old lambs 56.5–69.7 % FECR40–53.5%% WB reduction


Smith et al. ,1999

H-gal-GP 100µg Quil A

Lambs 83.4% WB reduction


Newlands et al.., 1999

H-gal-GP 100 µg Quil A

Lambs 93% FECR 60–64%WB reduction


Smith et al. ,2000Smith et al. ,2003Smith et al. ,2007

H-gal-GP 100 µg Quil A

Lambs 88.5% FECR72.3% WB reduction


Cachat et al. ,2010

Vaccination with native H-gal-GP

Location Animal specification

Antigen ,dose and adjuvant

Efficacy Reference

Lousiana,USA Suffolk ewes>2y 100 µg H11+100 µg H-gal-GP in 5 µg Quil A

>65%FECR Kabagambe et al., 2000

South Africa 12-18 m old Dorper sheep

100 µg H11+100 µg H-gal-GP in 5 µg Quil A

>82% FECR Smith et al., 2001

Australia Grazing merino lambs

100 µg H11+100 µg H-gal-GP in 5 µg Quil A

>85% FECRHigh levels of IgG1 and IgG2

LeJambre et al., 2008

Brazil Periparturient ewes 5 or 50 µg H11 and 1mg of saponin as adjuvant

78% FECR Bassetto et al., 2014

Field trials in sheep

1st commercially available sub-unit vaccine against H. contortus developed by Dr. David Smith of Moredun Research Institute, Edinburgh , UK.

Manufactured by Dept. of Agriculture and Food, Albany, Western Australia

5µg native gut integral protein + 1mg saponin adjuvant /dose (1ml Subcutaneously)

5 doses for lambs during the summer Haemonchus risk period

Reduces the periparturient rise – epidemiological benefit to flock

Thank you

vaccination against haemonchus contortus

Science