Nathalie Ospina - Daniela Ortiz

1. NATHALIE OSPINA ORTEGA
DANIELA ORTIZ PIEDRAHITA

2. INTRODUCTION
Brucella abortus
- intracellular bacteria
- Gram negative
- found in cattle populations
- Zoonotic
- Blood borne pathogen
- Causes premature abortion Of cattle fetus
- Causes brucellosis
Brucellosis
- signs and symptoms:
- Fever, joint pain, fatigue
- Treated with antibiotics
- Infection can recur
- No human vaccine
- killed vaccines for animal brucellosis
causes disadvantages

3. INTRODUCTION
immunization
- Public Health achievement
- Active: inoculation of microbial agents
- Passive: transfer of antibodies preformed
- Brucella: mucosal vaccines → l.casei
- Vectored bacterium or virus that express
exogenous antigens
lactobacillus casei
- Gram positive Lactic acid bacteria as
antigen delivery system
- Promising for mucosal immunization
- found in mouth, colon, female genital
tract
- Used for diverticulitis, diarrhea,
infections and dairy Food production

4. GENERAL OBJECTIVE
The aim of this study was to design a mucosal vector vaccine against
brucellosis on the basis of OMP19 antigen using Lactobacillus casei
bacterium as a vector, and evaluate the immunogenicity of this
vaccine and its protective effect against challenges with virulent
B.abortus 544 strain.

5. METHODS: PCR
Polymerase chain reaction
⎻ the objective of this technique is the amplification of genes or dna fragment or indirectly
through rna, present in diverse mixtures from different sources.
⎻ is based on enzymatic in vitro amplification (geometrical increase in the number of a particular
dna sequence)
⎻ this method can be used to detect tumor cells, infectious microorganisms and many others
⎻ this method was used in this article in order to achieve a vector with a protein that could be
anchored to the cell wall and being exposed to the target

6. ⎻ IT INVOLVES VOLUNTARILY CREATING DNA MOLECULES MADE UP OF DNA FRAGMENTS FROM DIFFERENT
SOURCES.
⎻ RECOMBINING REQUIRES CUTTING AND THEN SPECIFICALLY JOINING THE DNA. THIS GENETIC
MANIPULATION WAS CARRIED OUT IN LARGE PART USING ENZYMES THAT ACT ON DNA SUCH AS RESTRICTION
ENDONUCLEASES AND LIGASES.
⎻ THIS METHOD WAS USED TO PRODUCE THE PT1NX-OMP19 RECOMBINANT PLASMID, USING THE AMPLIFIED
GENE PIECE AND DIGESTING IT WITH SUITABLE RESTRICTION ENZYMES.
METHODS:recombinant dna

7. METHODS:elisa
⎻ Enzymes are fixed by conjugation with compounds that have two reactive sites, one for an
antibody and one for the enzyme.
⎻ The enzymes used are: alkaline phosphatase and horseradish peroxidase
⎻ This method was used in the article to identify the specific anti-OMP19 IgA and IgG antibodies

8. METHODS:immunofluorescence
- IS AN ASSAY WHICH IS USED PRIMARILY ON BIOLOGICAL SAMPLES AND IS CLASSICALLY DEFINED AS A
PROCEDURE TO DETECT ANTIGENS IN CELLULAR CONTEXTS USING ANTIBODIES.
- IT USES FLUORESCENTLY LABELED SECONDARY ANTIBODIES TO VISUALIZE PROTEINS IN CELLS AND TISSUES,
AND CAN PROVIDE INFORMATION ABOUT THE TISSUE DISTRIBUTION OF A GIVEN PROTEIN AS WELL AS ITS
SUBCELLULAR DISTRIBUTION.
- THIS METHOD WAS USED TO DEMONSTRATE THE EXPRESSION OF OMP19 PROTEIN ON THE SURFACE OF
RECOMBINANT L. CASEI BACTERIUM.

9. RESULTS

10. RESULTS

11. RESULTS

12. RESULTS

13. AUTHOR AFFIRMATION AGREES / DISAGREES
Farahi et al he investigated the capacity of brucella
abortus. omp19 to elicit splenic
lymphocytes stimulation response
Pasquevich et al he evaluated the potential of brucella
abortus OMP19 as a suitable subunit
vaccine against brucellosis
Ribeiro LA et al he constructed a L. lactis strain
producing recombinant B. abortus
L7/L12 antigen as a first step towards
food-grade live vaccines against
brucellosis
DISCUSSION

14. CONCLUSIONS
- brucella infections generally happen via mucosal surfaces, therefore, the expansion of mucosal
administered vaccines could be beneficial to controlling brucellosis at pathogen entry location
- In lactic acid bacteria, probiotic Lactobacillus casei is considered as a promising antigen delivery
system for mucosal immunization, and it could help with others illnesses.
- Oral immunization with recombinant probiotic Lactobacillus casei vector vaccine expressing
OMP19 stimulates the immunity of mucosal and systemic, hence the fact that the vaccine produced
in this research could be a very good candidate for the development of a vaccine against human
and animal brucellosis.

15. CONCEPTUAL MAP: NATHALIE OSPINA

16. CONCEPTUAL MAP: DANIELA ORTIZ