Project Abstract

<p> Sequel to the earlier works by Omenyi et al which established the role of surface thermodynamics in various biological processes from the electrostatic repulsion and van der Waals attraction mechanisms, HIV-blood interactions were modeled. This involved the use of the Hamaker coefficient approach as a thermodynamic tool in determining the interaction processes. It therefore became necessary to apply the Lifshitz derivation for van der Waals forces as an alternative to the contact angle approach which has been widely used in other biological systems. The methodology involved taking blood samples from twenty HIV-infected persons and from twenty uninfected persons for absorbance measurement using Ultraviolet Visible Spectrophotometer (Ultrospec3100pro). From the absorbance data various variables (e.g. dielectric constant, etc) required for computations with the Lifshitz formula were derived. CD4 counts using the digital CD4 counter were also obtained. Due to the very large body of data involved, MATLAB software tools were employed in solving the ensuing mathematics. The Hamaker constants A11, A22, A33 and the combined Hamaker coefficients A132 were obtained using the values of the dielectric constant together with the Lifshitz equation. The harmonized Hamaker coefficients A132har and the absolute combined Hamaker coefficient, A132abs (an integral of all the values of the various Hamaker coefficients) for the infected blood samples were then calculated. The value of A132abs = 0.2587×10-21Joule (i.e. 0.2587×10-14erg) was obtained for HIVinfected blood. This value lies within the range of values derived by various researchers for other biological systems. Another significance of this result is the positive sense of the absolute combined Hamaker coefficient which implies net positive van der Waals forces indicating an attraction between the virus and the lymphocyte. This in effect suggests that infection has occurred thus confirming the role of this principle in HIV-blood interactions. A lower value of A131abs = 0.1026×10-21 Joule obtained for the uninfected blood samples is also an indicator that a zero or negative absolute combined Hamaker coefficient is attainable. A mathematical model for the HIV-blood interaction mechanism was developed from the principle of particle-particle interaction mechanism. To propose a solution to HIV infection, it is necessary to find a way to render the absolute combined Hamaker coefficient A132abs negative. As a first step to this, a mathematical derivation for A33 ≥ 0.9763×10-21Joule which satisfies this condition for a negative A132abs was obtained. To achieve the condition of the stated A33 above with possible additive(s) in form of drugs to the serum as the intervening medium will be the next step. This forms part of the suggested areas for further research. <br></p>

Project Overview

<p> </p><p>INTRODUCTION</p><p>1.1 Rationale:<br>At the 2001 Special Session of the UN General Assembly on AIDS, 189 nations agreed that AIDS was a national and international development issue of the highest priority [1]. Between December 2005 and March 2006, UNAIDS compiled data from reports obtained from 126 countries on HIV/AIDS prevalence. In sub-Saharan Africa a mature epidemic continues to ravage beyond limits that many experts believed impossible. Also, relatively new but rapidly growing epidemics in regions such as Eastern Europe and South-East Asia that may come to rival that of sub-Saharan Africa in scope, had erupted [2].<br>Over time diverse clinical approaches to the issue of HIV/AIDS have been employed to seek to proffer possible solutions to the threat. Progress in this regard has been slow and far in between but has given birth to some palliative measures which include the introduction of the Highly Active Anti-retroviral Therapy (HAART). However, the results have not actually shown an easy and comprehensive solution due to the rapid mutative genetic nature of the virus [3].<br>Much research has been and is still on, on this subject with a cure not yet in view. The choice to approach it via the vehicle of surface thermodynamics against the conventional clinical methods is a novel one. The optimism stems from the great successes recorded with this approach in related areas of biology and medicine. The role of surface properties in various biological processes is now well established. In particular, interfacial tensions have been shown to play an important, if not crucial role in phenomena as diverse as the critical closing and opening of vessels in the microcirculation, cell adhesion, protein adsorption, antigen-antibody interactions, and phagocytosis [4].</p><p>1.2 Background to Study:<br>The HIV is assumed to be a particle which is dispersed in a liquid (the serum) and attacks another particle (the lymphocytes). The virus attaches itself on the surface of the blood cell before penetrating it to attack the RNA. If the surface of the blood cell is such that it will repel the virus, access to the virus into the interior of the cell would have been denied. Thus, the initial actions take place on the surfaces of the cell and of the virus (assumed to be particles). This interaction which involves two surfaces coming together in the first instance can be viewed as a surface effect.<br>It therefore stands to reason that, if it is possible to determine the surface properties of the interacting particles, then one can predict the mechanisms of their interactions.<br>4</p><p>When two particles make contact, they establish a common area of contact. Some original area of the surface of each particle has been displaced, and the work done to displace a unit area of the surface is referred to as the surface free energy. The actions therefore that take place on the surfaces are termed surface thermodynamic effects. These actions are assumed to occur slowly so that thermodynamic equilibrium is assured. This concept will be employed in this research work to characterize the HIVblood interactions with the serum as the intervening medium.<br>The clinicians have analyzed the surfaces of blood cells on which the virus binds. There are receptors and coreceptors on these cells and suggested types and their roles in the attachment processes are given in tables 1.1 and 1.2. Figures 1.1 and 1.2 show the nature and the interactions of these cells.</p><p>Fig.1.1: Human Immunodeficiency Virus (HIV) Anatomy [5]<br>HIV infects immune cells by interacting with proteins on the cells’ surfaces. The CCR5 is the preferred co-receptor for HIV in the human immune system. Immune cells that express CCR5 respond to sites of injury or inflammation. In order to respond effectively, they must go to the site of action. When a tissue experiences trauma or inflammation, nearby cells secret signal molecules called chemokines. The chemokines diffuse out from the site of the trauma through the blood stream where they come in contact with cells expressing the appropriate receptor. Each cell expresses several different receptors so they can respond to different immune signals.<br>5</p><p>Figure 15b The<br>Fig.1.2: Interaction of a Dendritic Cell (right) having HIV bound to its surface (arrow) with a Lymphocyte (left) [6]</p><p>CCR5 is a seven trans-membrane protein or 7TM which means that it crosses the plasma membrane of the cell seven times. 7TM proteins are sensors for the cell. They communicate what happens outside the cell to the inside of the cell through a process called alosterism. The Chemokines bind to CCR5 which causes the CCR5 to change shape both outside and inside of the cell. The altered shape of CCR5 changes the interactions with G-proteins inside the cell initiating a signal transduction cascade that activates the cell to go to the site of injury.<br>The redundancy inherent in the immune system allows many Chemokines to signal for multiple coreceptors. CCR5 binds the Chemokine’s RANTES, MIP-1α and MIP-1β. It is important to note that these Chemokines also bind to other receptors. Both RANTES and MIP-1α can bind to CCR1 and RANTES can also bind to CCR3. This is an example of redundancy which is common in the immune system. In this way, if one pathway is blocked, the immune response can be achieved through another. Thus, as these receptors interact with the stream of Chemokines they direct the cell to the site of injury or inflammation. In summary, CCR5 plays an important role in the movement of immune cells to the site of action. The key points include;<br>ï‚· CCR5 is a censor protein on certain immune cells. ï‚· CCR5 binds to selected Chemokines like MIP-1α, MIP-1β and RANTES. ï‚· CCR5 transduces signals inside the immune cell. ï‚· These signals result in chemotaxis or movement of the cell to the site of<br>injury.</p><p>6</p><p>Table 1.1: Cell Surface Receptors Implicated in Binding HIV Virions: Receptors<br>other than CD4 or Coreceptors that attach HIV Virions to Cell Surfaces [7]<br>Receptor Affinity<br>(Kd)<br>Expression Role in attachment and<br>infection<br>Reference<br>Gal-C High<br>(11.6 nM)<br>Neuronal and<br>glial cells<br>Confers inefficient<br>infection presumably by<br>aiding attachment<br>Harouse et al. (1991)<br>Sulphatide (sulphate<br>derivative of Gal-C)<br>Colorectal<br>epithelial cells<br>and primary<br>macrophages<br>Confers efficient CD4<br>independent infection by<br>NDK, a TCLA HIV-1<br>strain Requires CXCR4<br>coreceptor<br>Fantini et al. (1993);<br>Seddiki et al. (1994);<br>Delezay et al. (1997)<br>Placental<br>membrane-binding<br>protein<br>High<br>(1.3–0.6<br>nM)<br>Cloned from a<br>placental<br>cDNA library<br>Binds virus particles to<br>the cell surface and thus<br>enhances infectivity via<br>CD4 and coreceptors.<br>May trap HIV in the<br>periphery and carry to<br>T-cells in lymph nodes<br>Curtis et al. (1992);<br>Geijtenbeek et al.<br>(2000)<br>DC-SIGN On dendritic<br>cells</p><p>DC-SIGNR</p><p>Endothelial<br>cells, such as<br>liver,sinusoidal<br>and lymph<br>node sinus<br>endothelial<br>cells<br>Acts in the same way as<br>DC-SIGN<br>Pohlmann et al.<br>(2001)<br>Mannose-specific Macrophages Binds gp120 Larkin et al. (1989)<br>7</p><p>macrophage<br>endocytosis receptor<br>Heparans</p><p>Many cell<br>types<br>Attaches virus particles<br>to cell surfaces via an<br>interaction with the V3<br>loop thus enhancing<br>infectivity via CD4 and<br>coreceptors. Acts<br>predominantly for<br>CXCR4-using viruses<br>Mondor et al. (1998)<br>LFA-1/ICAM-1</p><p>LFA-1 is<br>expressed on<br>haematopoietic<br>cells, ICAM-1<br>is on a wide<br>variety of cell<br>types<br>ICAM-1 encorporated<br>onto virions enhances<br>attachment and infection of LFA-1+ cells<br>Fortin et al. (1999);<br>Paquette et al.<br>(1998)</p><p>8</p><p>Table 1.2: Human Polymorphisms in Chemokine and Coreceptor Receptor<br>Genes that influence HIV Infection and Disease Progression [7]<br>Genotype<br>Frequency<br>Effect<br>CCR5 32/wild-type Up to 18 % in<br>Caucasians<br>Slows disease progression<br>CCR5 32/ 32 Up to 1 % in<br>Caucasians<br>Protects against infection<br>CCR5 m303 leads to premature stop<br>codon and CCR5 truncated in E1<br>3/209 healthy<br>donors<br>In combination with a 32 CCR5 allele confers<br>T-cells with resistance to R5 viruses<br>CCR5 P1 allele, characterized by a pattern<br>of 10 specific bases at different sites,<br>including A at –2459<br>43–68 % Accelerates disease progression<br>CCR5 A/G at –2459 43–68 % Slows disease progression<br>CCR2 V64I is linked to a point mutation in<br>the promoter region of CCR5<br>10–15 % in<br>Caucasians and<br>US Africans<br>Slows disease progression<br>SDF-1 in 3´ untranslated region of mRNA.<br>In SDF-1 but not SDF-1 mRNA<br>16–25 % Homozygotes have slower disease progression, even slower if 32/wild-type CCR5 or V64I<br>CCR2 also<br>RANTES promoter AC, GC and AG at<br>sites –471, –96 (sites equivalent to –403<br>and –28 as described by Liu et al., 1999)<br>Variable<br>depending on<br>population<br>Faster/slower disease progression depending on<br>genotype and population (Gonzalez et al., 2001).<br>Some protection from transmission if –471A<br>present<br>MIP-1 intron +113, +459 Variable<br>depending on<br>population<br>Faster/slower disease progression depending on<br>genotype and population (Gonzalez et al., 2001)</p><p>9</p><p>1.3 Statement of Problem:<br>The discovery and application of highly active anti-retroviral therapy (HAART) to suppress HIV has revolutionized the clinical management of HIV/AIDS cases. The HIV however, has the capacity to develop resistance to the antiretroviral drugs and this phenomenon has turned out to be a significant cause of failure of HAART. HIV, being an RNA-based rapidly mutating virus, (unlike the DNA-based counterparts) lacks the ability to check for and correct genetic mutations that can occur during replication. In chronic HIV cases, about ten billion new viral species can be generated daily. This rapid genetic variation has made it rather very difficult to proffer a clinical solution to the problem [3] and the worldwide picture is one of increasing rates of infection [8].<br>It is against this backdrop that this study explores a novel and rare approach using surface thermodynamics to seek a way forward in the research on the topic of HIV-blood interactions. The successes recorded in the use of this approach in finding solutions that have brought about many scientific applications cannot be overemphasized [4].</p><p>1.4 Objective of the Study:<br>This research work is aimed at employing the concept of surface thermodynamics to study the interaction between the virus and the blood cells with a view to proffering a solution to the HIV/AIDS pandemic. The following tasks therefore, must be kept in view;<br>(i) Determine the mechanism of interaction of HIV with white blood cells.<br>(ii) Seek a thermodynamic interpretation of such interactions through van<br>der Waals attraction mechanism.<br>(iii) Quantify such interactions through actual measurements.<br>(iv) Recommend possible approach to eliminating the HIV-blood<br>interactions.<br>Thus, the main thrust of this research work is the use of surface thermodynamics in explaining the HIV/AIDS jinx.</p><p>1.5 Scope and Limits of the Study:<br>The scope of this research is limited to specifying the relevance of van der Waals forces to the fusion of the HIV with the receptor cells and how such fusion process could be quantified and prevented. This is intended to be achieved by the application of surface thermodynamics using the concept of Hamaker coefficients derived from absorbance data required for the computation of the Lifshitz formula. The sign of the<br>10</p><p>combined Hamaker coefficient will suggest whether there is attraction or repulsion between the virus and the blood cells.<br>The next approach would be to suggest a formulation that would aid in preventing contact between the virus and the blood cell, and hence prevent their interactions. This entails the development a model that would render the absolute combined Hamaker coefficient, A132abs negative thus causing the virus and the lymphocytes to repel each other [9].<br>Sourcing for additives to achieve this aim is beyond the scope of this work. Other approach for the determination of Hamaker coefficients, e.g. by contact angle data, will not be considered. These will serve as suggested areas for further research.</p> <br><p></p>