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CHAPTER 4 AN INTRODUCTION TO CELL STRUCTURE AND HOSTPATHOGEN RELATIONSHIPS © National Institutes of Health, United States Department of Health and Human Service. WHY IS THIS IMPORTANT? • An understanding of cell structure is vital to understanding microbiology, in particular, the process of infection. • Host-pathogen interactions must be understood in order to understand the processes of infection and disease. OVERVIEW CLASSIFICATION OF ORGANISMS • All living organisms can be classified as Prokaryotes Eukaryotes No Organelles Organelles (Bacteria) (Everything else) Biological Kingdoms Taxonomy: A system for classifying biological organisms Genus and species Used to identify an organism CLASSIFICATION OF ORGANISMS • Biologists classify microorganisms by their genus and species names. – An example is Clostridium tetani. Clostridium is the genus and tetani is the species. • The genus can have several species. – E.g. Clostridium perfringens, Clostridium botulinum and Clostridium difficile, etc. • The genus and species names of microorganisms are italicized when written. • First letter of genus is capitalized • The genus can be abbreviated to first letter: Escherichia coli E. coli All E’s are NOT Created Equal The genus can be abbreviated to first letter: Escherichia coli E. coli Enterobacter aerogenes Enterococcus faecalis Staphylococcus aureus Streptococcus pneumoniae E. aerogenes E. faecalis S. aureus S. pneumoniae BACTERIA: Overview • Microscopic • Immensely diverse • Successfully colonize all parts of the world and its inhabitants. • Considered Pathogens if they cause disease. BACTERIA: Size, Shape and Multicellular Arrangement • Bacteria can be of different shapes, sizes, and arrangements. • The most common shapes: – Coccus (circular) – Bacillus (rod) – Spirilla (spiral) Multicellular Arrangements Chain Clusters, clumps Many organisms have no arrangement. BACTERIAL STAINING: Types of Stain • Bacteria can be stained in the following ways: – – – – Positive stains – stain the organism Negative stains – stain the background Simple stains – stain using only one color Differential stains – stain using more than one color THE GRAM STAIN • Differentiates bacteria based on differences in the structure of their cell walls. • The Gram stain process divides bacteria into four major groups: – – – – Gram-positive Gram-negative Gram-variable Gram nonreactive Gram neg. bacilli Gram pos. cocci THE GRAM STAIN THE (CAPSULE) STAIN • Stains theNEGATIVE background surrounding encapsulated bacteria. • Capsules serve as important virulence factors for some organisms: • Streptococcus pneumoniae (pneumonia, meningitis, etc.) • Hemophilus influenzae (pneumonia, meningitis, etc.) • Neisseria meningitidis (bacterial meningitis) • Bacillus anthracis (anthrax) THE FLAGELLA STAIN • The flagella stain identifies the presence of flagella, which are used for motility. • Motility is important for infection as it allows the invading organisms to move from the initial site of infection. THE ZIEHL-NEELSEN ACID FAST STAIN • Used to detect Mycobacterium species such as M. tuberculosis (the cause of tuberculosis) or M. leprae (the cause of leprosy). • These organisms have mycolic acid in their cell walls, making the cell wall difficult to penetrate. THE ENDOSPORE STAIN • Endospores are small, tough, dormant structures that can form in certain bacteria. Crystal Violet Stain Excluded from spores Malachite Green Spore Stain HOST-PATHOGEN RELATIONSHIPS • Infectious diseases are complex and involve the ongoing interaction between the host and the pathogen. – HOST PATHOGEN • Many factors affect this relationship. HOST-PATHOGEN RELATIONSHIPS • Infectious diseases are complex and involve a series of shifting interactions between host and pathogen. • For the pathogen, the interactions depend on: – Ability to evade or overcome the host’s defense – Ability to increase in numbers – Ability to transmit to new hosts. Covered in greater detail next week. HOST-PATHOGEN RELATIONSHIPS • For the host, the interactions depend on: – The host having useful functioning defenses – The host’s susceptibility to infection Covered in greater detail in weeks 6 and 7. PATHOGENICITY: All Microbes Are Not Created Equal Commensals, Opportunists and Pathogens Commensals – Do not harm the host Mutualists – Benefit the host while deriving benefit from the host Opportunists – Can become pathogens given the proper conditions (injury, immune compromised, etc.) Pathogens – Overtly cause disease HOST-PATHOGEN RELATIONSHIPS • Microbial flora can protect us through microbial antagonism. – Many bacteria produce bacteriocins which are localized bacterial antibiotics. – Bacteriocins can kill invading organisms but do not affect the bacteria that produce them. Competition for space and other resources. • In most cases, the interactions between the body and bacteria cause no harm. – Some bacteria have a mutualistic relationship with the host. • However, some harmless organisms can become opportunistically pathogenic. OPPORTUNISTIC PATHOGENS AND PRIMARY PATHOGENS • Opportunistic pathogens cause infection by taking advantage of a hosts’ increased susceptibility of infection. • Characteristics of primary pathogens are as follows: – They cause disease in healthy individuals. – They include viruses and bacteria. OPPORTUNISTIC PATHOGENS AND PRIMARY PATHOGENS • Characteristics of primary pathogens are as follows: – They have evolved mechanisms that can overcome host defenses. – Once inside, they can multiply rapidly. – Some primary pathogens are restricted to humans. – Some pathogens can infect humans and other animals (zoonosis) DISEASE AND TRANSMISSIBILITY • Successful infection requires the following from a pathogen: – The ability to multiply in sufficient numbers – The ability to transmit to new hosts. DISEASE AND TRANSMISSIBILITY • Symptoms of certain infections can provide transmission mechanisms through: – Coughing transmits respiratory infections. – Diarrhea transmits digestive infections. • Infections that kill too quickly inhibit transmission. BACTERIAL PATHOGENICITY AND VIRULENCE • Pathogens must be able to accomplish the five requirements for infection: – – – – – Entry (getting in) Establishment (staying in) Defeat the host defenses Damage the host Be transmissible •Entry •Colonization •Immune Evasion •Propagation •Transmission PATHOGENICITY AND VIRULENCE • Virulence refers to how harmful a pathogen is to the host. – Virulence depends on genetic factors of the pathogen. – These genetic elements are often turned on only in the host. BACTERIAL PATHOGENICITY AND VIRULENCE • Pathogens carry virulence genes in clusters called pathogenicity islands. – These can be located on plasmids. – Plasmids can be transferred between cells. QUORUM SENSING • Organisms sense their environment using special sensing proteins. This is called quorum sensing. • This sensing is based on population densities. • Certain genes are only turned on when there are enough cells present: An example of this is enterotoxin production in Salmonella. BIOFILMS • Bacteria can grow in aggregated assemblies called biofilms. • Biofilms are clinically important because: – They can capture and retain nutrients (allowing continued growth). – They impede uptake of antibiotics and disinfectants. – They inhibit phagocytosis. BIOFILMS • Biofilms can build up on medical devices such as: – Catheters – Heart valves – Prosthetic devices Lead to nosocomial (hospital acquired) infections • Biofilms are one of the causes of plaque buildup on teeth. THE HOST CELL • There are several differences between prokaryotic and eukaryotic cells. • Animals, including humans, possess eukaryotic cells. • Many of the structures of eukaryotic cells play a role in infection. EUKARYOTIC CELLS: Bacterial anatomy will be covered in week 3 (chapter 9). EUKARYOTIC CELLS: The Plasma Membrane • The eukaryotic plasma membrane is made up of a phospholipid bilayer. • It is a fluid matrix containing a variety of proteins and other molecules. EUKARYOTIC CELLS: Role of the Plasma Membrane in Infection • Because the plasma membrane is the barrier between the inside and the outside of the cell, it must be breached if pathogens are to gain entrance. EUKARYOTIC CELLS: Cytoplasm • The eukaryotic cytoplasm consists of: – A semi-fluid material that is mainly water and dissolved substances – Membrane-bound structures called organelles – Structures not bound by membranes. EUKARYOTIC CELLS: Role of the Cytoplasm in Infection • Cytoplasm is involved in a variety of infections. • It has a major role in viral infections. • Many viruses replicate in the host cell cytoplasm. EUKARYOTIC CELLS: Host Cell Cytoskeleton • The cytoskeleton gives eukaryotic cells structural integrity. • The cytoskeleton is involved in how cells are joined together to form tissue. • The components of the cytoskeleton play a role in cellular mitosis and meiosis. Role of the Cytoskeleton in Infection • Many pathogens use the cytoskeleton as part of the infection process. • Shigella use microfilaments to move laterally between cells of the intestine. EUKARYOTIC CELLS: Cilia • Cilia are made up of microtubules that can be projected outward from the cell surface. • The lower respiratory tract which is lined with ciliated cells that work together with mucusproducing cells to move trapped particles upward and out of the respiratory tract. • Muco-cilliary elevator EUKARYOTIC CELLS: Role of Cilia in Infection • Pathogens can attack the cilia and destroy their trapping capability. • In some respiratory diseases, such as pertussis (whooping cough), the pathogens (in this case Bordetella pertussis) attach to host ciliated cells as an initial part of the infection. EUKARYOTIC CELLS: Ribosomes • Ribosomes are the site of protein synthesis. • They are found either individually or attached to the endoplasmic reticulum. • Ribosomes in eukaryotic cells have a different structure to those prokaryotic cells. – Difference used to develop antibiotics EUKARYOTIC CELLS: Role of the Ribosome in Infection • Eukaryotic ribosomes are very important in viral infections. – The virus takes over the host cell ribosome function. – It is then used only to make new virus. ER & Golgi Apparatus • Both systems of membranes that form flattened sacs. • The endoplasmic reticulum (ER) can be smooth (without ribosomes) for lipid synthesis, or rough (with attached ribosomes) for protein synthesis. ER & Golgi Apparatus • The Golgi apparatus has the three following functions: – Modifying and packaging products coming from the ER – Renewing the cell’s plasma membrane – Producing lysosomes The ER & Golgi Apparatus in Infection • Both structures are involved in the biosynthesis and assembly of viruses. • The ER is also involved in the adaptive immune response to infection. Lysosomes • Lysosomes are filled with destructive enzymes and chemicals. • They destroy foreign materials that enter the cell. • They also act in recycling host cell components. Role of the Lysosome in Infection • Lysosomes fuse with phagocytic vesicles and destroy invading pathogens. • Many pathogens can defeat this defense. EUKARYOTIC CELLS: The Nucleus • The nucleus is the location of the cellular DNA of eukaryotic cells. • The nucleus is bound by a double phospholipid bilayer membrane. • It is the site for DNA replication during cell division. • The transcription of messenger RNA also occurs here. EUKARYOTIC CELLS: Role of the Nucleus in Infection • The nucleus of the host cell is important in many infections, particularly those caused by DNA viruses. – Copies of the viral DNA are made in the nucleus. EUKARYOTIC CELLS: Endocytosis & Exocytosis • Endocytosis involves bringing things into the cell through the formation of vesicles. • Exocytosis involves moving things out of the cell which is also done through the formation of vesicles. Endocytosis & Exocytosis • There are three ways that endocytosis operates within a cell: – Pinocytosis – Phagocytosis – Receptormediated endocytosis Endocytosis & Exocytosis in Infection • Many pathogens enter the host cell through the formation of vesicles. • This method provides protection for the pathogen from the host immune response. • Some pathogens bind to host cell receptors that trigger endocytosis. This is particularly true of viruses. EUKARYOTIC CELLS: Role of Endocytosis & Exocytosis in Infection • Phagocytosis is a type of endocytosis that can be used to defend against infection. • Many pathogens have found ways to defeat phagocytosis.