Climate Change – An Urgent Call to Action

Humanity does not seem to grasp the dim future that awaits the entire species if the dire warnings issued by climate scientists worldwide are not heeded regarding the absolute necessity to significantly diminish the burning of fossil fuels and thereby slow down the inexorable increase of the concentration of greenhouse gases in the atmosphere.

Ironically, human families work diligently towards fashioning a better life for their children, grandchildren and future descendants, yet their focus has been tilted towards material success and enhanced prosperity.  These are important goals.  However, if this onward rush towards greater and greater prosperity is not tempered by reality, the insidious consequences of climate change will make a significant portion of the planet essentially uninhabitable and rising seas will inundate the habitats of hundreds of millions of human populations wreaking havoc upon centers of economic power where great stores of human wealth reside.  Under such circumstances, such wealth would immediately be rendered quite meaningless.

It is quite absurd at this juncture to argue the merits of the science or allow short-term economic interests to resist the necessity for immediate and significant action.  If we do not act quickly as a world community, our descendants will find themselves marooned on what would increasingly become a hostile planet.  Is this the kind of future we are working so assiduously towards?

Obfuscation and resistance serves no useful purpose – the longer we procrastinate the more impossible it will be to make meaningful corrections on this our spaceship earth.

Ultimately, the science speaks for itself. 

Use of Genetically Modified Yeast in the Production of Opiates

Opiates are pain-relieving drugs that play a vital role in palliative care.  Morphine is a well-known member of this class of compounds.  Opiates have been derived from poppy plants for thousands of years.  The compound thebaine (paramorphine or codeine methyl enol ether) - see structure below -is the natural product that is the precursor for opiates.

Thebaine

Due to the incredible strides that have been made in regards to genetic engineering, a team of researchers headed by Christina Smolke from Stanford University in Palo Alto California has successfully incorporated genes derived from a variety of plants including poppies, bacterial genes from Pseudomonas and rodent genes into the yeast genome and endowed these modified yeast organisms with the capacity to produce thebaine from a sugar source.  This methodology required the successful incorporation of 21 different genes – a monumental enterprise.

This is a significant breakthrough in the area of synthetic biology for it suggests that eventually it will become possible to readily produce large quantities of thebaine that can subsequently be used to produce a variety of opiates.  Smolke and her colleagues avoided producing a modified yeast cell capable of producing morphine directly from sugar in order to forestall the possibility of inadvertently making the illicit production of morphine a practical reality.

The Role of Neutrophils in Guiding the Immune Response to Infection by the Influenza Virus

The influenza is virus has a global impact on human populations. Much research effort has been directed towards understanding how the human immune system combats influenza virus infection. The cellular target of this virus is the epithelial cells of the respiratory tract. In order for the immune system to successfully combat this pathogen, virus specific cytotoxic CD8+ circulating lymphocytes must migrate to the site of infection.

 It has been established that in order for these cytotoxic CD8+ lymphocytes to successfully kill virally-infected cells, it must be preceded by the innate immune response. A key player in this first response are Neutrophils (see image below) These cells are generally the first cell type to cross the blood vessel epithelium into the distressed tissue and are responsible for generating chemical signals that alert different types of immune cells.

 Although this relationship between Neutrophils and aspects of the adaptive immune response has been established, the underlying molecular mechanisms have been unclear. Dr. Kihong Lim and his colleagues at the Department of Microbiology and Immunology, David H. Smith Center for Vaccine Biology and Immunology, University of Rochester, Rochester, NY have investigated the mechanism of this immune response to infection by the influenza virus. What they uncovered was an interesting process.

Their studies have shown that the subset of cytotoxic CD8+ lymphocytes specific for influenza infected epithelium cells is dependent upon the presence of Neutrophils at the site of infection. The migrating Neutrophils leave in their wake a chemical trail; that chemical is a particular chemokine – a class of substances that serves as a powerful attractant to circulating lymphocytes – CXCL12. It is this CXCL12 that is a necessary requirement for the procurement of virus specific CD8+ lymphocytes at the infection site and for their subsequent ability to kill infected respiratory epithelial cells.

The results of this work contribute significantly to the fundamental understanding of the complex interactions that are required to mount a successful immunological assault upon a virus-induced infection.

Progress in Developing a Vaccine against the Ebola Virus

Progress in Developing a Vaccine against the Ebola Virus

The recent spread of hemorrhagic fever (EHF) in areas of West Africa including Guinea, Sierra Leone and Liberia has placed considerable urgency on the need to develop an effective vaccine against the pathogen responsible for this horrific and highly contagious disease, the Ebola virus (EBOV).  To date, of the approximately 27,200 reported cases, there have been more than 11,100 deaths – a mortality rate of  40.8 percent.   EBOV is so infectious that a high incidence rate has also been reported among health care workers.  It needs to be kept in mind that the health care infrastructure of the countries affected has been severely compromised especially since these so-called, “low resource” countries have budgets inadequate to respond effectively to this challenge.

By way of background EBOV belongs to a class of viruses called filoviruses.   Filoviruses are single-stranded RNA viruses.  This class of viruses possesses a glycoprotein (GP) on its surface that could conceivably make a good candidate as an immunogen – a protein capable of eliciting an immune response.

Dr. Andrea Marzi at the Laboratory of Virology, Division of Intramural Research at the National Institute of Allergy and Infectious Diseases  at the National Institutes of Health in Hamilton Montana.and his colleagues  have helped develop a strategy for developing such a vaccine.  The strategy they employed can be outlined in the following way –

• A live attenuated stomatitis virus was employed as a viral vector
• Recombinant technology was employed to modify this virus so that it expresses the Ebola GP on its surface referred to as Viral Stomatitis Vector EBOV (VSV-EBOV)
• This viral vector was then introduced into experimental animals – the rodent and macaque.

This approach was shown to be highly efficacious in both pre and post exposure vaccinations.  These results were so promising that phase 1 clinical trials in humans  were begun in several  worldwide locations. 

The following is in the author’s own words – “Complete and partial protection was achieved with a single dose given as late as 7 and 3 days before challenge, respectively.  This indicates that VSV-EBOV may protect humans against EBOV infections in West Africa with relatively short time to immunity, promoting its use for immediate public health responses.”

Tumor Suppressor p53 and the Clearance of Apoptotic Cells

Within the human body, with its complex array of highly differentiated organs and tissues, it is estimated that more than a billion cells die each day.  Most of these deaths arise from a natural process referred to as apoptosis – programmed cell death.  Since apoptosis is integral to the functioning of a healthy organism, its mechanism has been the focus of study for many years. 

In addition to the process of apoptosis, there also exists a mechanism for the efficient processing and clearance of the cellular debris that is a direct result of apoptosis.  Otherwise, the accumulation of released substances from dead cells could function as autoantigens and elicit an undesirable immune response against normal tissue resulting in autoimmune diseases.  In addition, chronic inflammation and developmental abnormalities could result from the buildup up cellular debris.  It has been established that the immune response to apoptotic cells involves the mobilization of phagocytic cells whose function is to engulf the dead or dying cells.  This process necessarily requires the expression of immune tolerance in order to prevent an autoimmune response.

The tumor suppressor  p53 protein has long been associated with the mechanism of apoptosis.  However, little has been established in regards to the putative role of p53 in the clearance of cellular debris that results from apoptosis.

Dr. Kyoung Wan Yoon and his colleagues from the Cutaneous Biology Research Center, Massachusetts General Hospital and Harvard Medical School, Charlestown, MA have focused their research efforts on the elucidation of the role of p53 in the clearance of post apoptotic cells and the establishment of tolerance to self-antigens.

What they have shown is the following.  As a result of stress upon a cell that leads to p53-mediated apoptosis, p53 triggers the production of a protein product from the so-called “Death Domain1a (DD1a)” gene.  DD1a functions as a ligand that is instrumental in the binding of the apoptotic cell to the phagocytic immune cell that will eventually engulf the dead cell.  Once this engulfment occurs it subsequently triggers the binding of an inhibitory T cell that ensures tolerance to self-antigens; thereby, preventing untoward reactions that could lead to a disease state.

This kind of work is extremely valuable in furthering the understanding of the mechanism of self-tolerance.  Such knowledge may prove instrumental in determining the underlying mechanisms involved in auto-immune disease.

HIV-1 Vaccine Development

It has been over thirty years since the discovery of HIV-1 as the virus responsible for AIDS.  It has been a long-standing goal to produce a vaccine able to induce cross-reactive antibodies that are capable of neutralizing the infectious capability of the HIV-1 virus in all its variant forms.  Progress in this direction has been impeded due to the complexity of the surface envelope glycoprotein (ENV) – a glycoprotein is a class of proteins containing carbohydrate moieties.  ENV is responsible for the process that allows the entry of the HIV-1 virus into its host cell (CD4⁺ T Helper cells).

ENV is trimer consisting of three glycoproteins with a molecular weight of 160K.  This trimer is split into a 120K surface component and a 41K membrane component.  Together they constitute a moiety that facilitates the entry of the virus into the host cell called the viral spike.  Although the components of the viral spike have long been considered candidates for a vaccine, it has been elusive.  In addition, the viral genes for these components have been discovered and used in vaccine production; this approach has also been shown to be unsatisfactory.  The reason for this failure seems to reside in the fact that many of the regions of the molecular structure of the ENV subunits that are candidates for eliciting a significant immune response lie buried in areas that are effectively hidden from immune-surveillance.

As a consequence, investigators have proposed using the entire ENV trimer as an immunogen – a substance capable of stimulating the immune system to produce humoral antibodies against it.  Unfortunately, the stability of ENV outside the environment of the viral membrane rapidly degrades.  However, this intrinsic difficult has been effectively sidestepped by R.W Sanders and associates (Science 349, aac4223(2015)).  They overcame this obstacle by engineering a molecule with covalent disulfide bonds that held the subunits together.  They subsequently used this modified ENV to immunize rabbits and monkeys.  Although it worked effectively against the same strain of virus from which the ENV was obtained, it was ineffective against heterologous strains of the deadly virus.

Although the problem of creating an effective vaccine against HIV-1 remains intractable, future efforts and innovative approaches similar to ones reported here may finally yield a viable solution.  Progress in science necessarily depends on all the work that has gone on before and the collaborative effort and energy of many. 

The role of T cells in Establishing Self-Tolerance

The human immune system is a powerful system designed to protect the individual from the onslaught of deleterious microorganisms that populate the natural environment.  Since it is essential that immune-capable cells not attack the tissues of the host, mechanisms for self-tolerance are necessarily implemented early in development.   When this self-tolerance mechanism fails, the result is often expressed as an autoimmune disease.  An example of such an ailment is multiple sclerosis (MS) in which the immune systems produces antibodies against myelin – the proteins that provides insulation for the electrical impulses that travel through the peripheral nerves.

Given the important role that self-tolerance plays in human health, many research laboratories are involved in fully elucidating its mechanism.   It is well known that the thymus gland is the site where self-tolerance is established.    It has also been shown that the immune regulator protein Aire is an important factor in the establishment of immunological tolerance; it operates within a subset of thymic stromal cells and directs T cell selection.  Aire is a transcription factor expressed in the medulla of the thymus and controls the mechanism that prevents the immune system from attacking the body itself.  Individuals with the autoimmune disease polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED) have been shown to have a mutation in the AIRE gene.

Since Aire seems to play such a fundamental role in the early development of immuno-tolerance, it would be of extreme interest to delineate the underlying molecular mechanism for this affect.  Dr. Siyoung Yang and his colleagues at the Division of Immunology in the Department of Microbiology and Immunobiology at Harvard Medical School in Boston have made some interesting discoveries in this regard.  They have reported that Aire promotes the creation of a distinct population of regulatory T cells – Foxp3⁺CD4⁺ - in the very early stage of development.   Furthermore, they found that these regulatory T cells persist into adulthood and play a pivotal role in self-tolerance.

This is very important that contributes significantly to the understanding immune-tolerance and this kind information may prove invaluable in the understanding and eventual treatment of autoimmune diseases.